Dijkstra, Klaas,

Hangyi, Ilona W.,

Kroon, Gerard J.A.,

Renken, Remco J.,

Robillard, George T.,

Scheek, Ruud M.,

Thole, Esther R.,

Hangyi, Ilona W.

Kroon, Gerard J.A.

Thole, Esther R.

Renken, Remco J.

Dijkstra, Klaas

Scheek, Ruud M.

Robillard, George T.

University of Groningen Research Database

  
 University of Groningen
Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by
Heteronuclear NMR Spectroscopy
Hangyi, Ilona W.; Kroon, Gerard J.A.; Thole, Esther R.; Renken, Remco; Dijkstra, Klaas;
Scheek, Ruud M.; Robillard, George T.
IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from
it. Please check the document version below.
Document Version
Publisher's PDF, also known as Version of record
Publication date:
1996
Link to publication in University of Groningen/UMCG research database
Citation for published version (APA):
Hangyi, I. W., Kroon, G. J. A., Thole, E. R., Renken, R. J., Dijkstra, K., Scheek, R. M., & Robillard, G. T.,
(1996). Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by Heteronuclear
NMR Spectroscopy, 1 p.
Copyright
Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the
author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).
Take-down policy
If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately
and investigate your claim.
Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the
number of authors shown on this cover page is limited to 10 maximum.
Download date: 11-02-2018
P-Al-09 
lUNAW.F%AHF@~J.A. 
KROON, ESTHWRTHOLE,REMCOJ. 
RENKEN,KLAASDUKS7RA, RUUDM. 
!$CHEEK and GEORGE T. ROBILLARD. 
GBB, University of &m&en (NL). 
l!auym-II mannitol (EIFll) of E coli is part 
ofa@emthatregulatestheuptakeand 
phosphorylation of specific carbohydrate 
subsbks.Ourgroupstudiesthe 
strutitiction relationship of the 
cytoplasmic domains of IZ.IF using 
multidimensional NMRtechniques. The G 
teminal domain of EV’ (ElIA”“‘, 148 
residnes, 16.4 kD) has heen cloned and 
ovtsqmsd in E cob. 
The low resoldon strwtwe was determined 
using 3D NMR techniqnes on ‘%I enriched 
protein and ‘SN/‘F labeled protein. ELV’ 
consists of a twwtranded psheet surrounded 
by 5 cchelices. 
Mutant ElIAd (ElIAH65Q) that cannot be 
phosphorylated by phospho-HPr (MIPr) was 
constructed. The bii interface beti 
EIIAJkl65Q and I-R is cow to that of 
EIIAH65Q and P-I-B, using 2D ‘%-HSQC. 
P-Al-l 1 
CRYSTAL STRUCTURE OF THE 13 
SUBUNIT HEART CYTOCHROME C 
OXIDASE. 
Shinzawa-Itoh, H,I Tsukihara, T,2 Aoyama, H.2 
Yamashita, E.2 Tomixaki, T.2 Yamaguchi, H,2 
Nakashima. R.1 Yaono, R.1 Yoshikawa , S.1 
1Himeji Institute of Technology (JP), 2Institute for 
Protein Research, Osaka University (JP). 
Purpose: For elucidation of the reaction 
mechanism of cytochrome oxidase, crystal 
structure of the enzyme was solved. 
Methods: The fully oxidized bovine heart 
cytochrome c oxidase stabilized with decyl 
maltoside was crystallized and the crystal 
structure was solved with MIR method at 2.8 A 
resolution with an R value of 20.4 % . 
Results and conclusion: Each structure of all 
the residues of the monomer (1780 in total) 
except for 23 residues has been converged to a 
reasonable structure by structural refinement. 
A hydrogen bond system including an imidazole 
bound to CuA. a peptide unit and a propionate 
of heme a could be an effective electron transfer 
path between CuA and heme a. Two structures 
spanned from the cytosolic surfaoe to the matrix 
including hydrogen bonds and internal cavities 
likely to contain water molecules, could serve as 
proton pumping path. Possible channels, for 
chemical protons to produce H20, for removing 
the produced H20 and for 02 were identified. 
Al 3D structures: experimental determination 
P-Al-10 
STR- OF MARE LiWTOF’E~ 
AT 4.0& RRSOLtiON 
KAUR P, SH&&A AK, KAR’lWKEYAN S, 
MITRA SN AND SINGH TP. 
Depmnt of Biophysics, AlI,lndia Institute of Medical 
Sciences, New Delhi-l 10 029 (India) 
Purpose: Lactofenin an iron binding glyco- 
protein (MW=80,OOODa) has two structural 
lobes, each housing one Fe’+ and a synergistic 
COs’- ion. Stucture determination has been 
carried out to understand the mechanism of 
action and the functional role played by 
lactofetin. 
Method: Purification from mare colostrum/milk 
by ion exchange and gel filteration. Crystalliza- 
tion by microdialysis method. structure 
elucidation using Molecular Replacement. 
Results:Tbe protein crystallizes in orthorhombic 
space group ~2~2~2, with a=79.8A, b=lOsSA, 
?=112.0A, 2=4 d an a solvent content of 57%. 
The structure has been refined for 6474 reflec- 
tions in the resolution range IO-4A. The current 
R factor is 0.27. The model contains 690 amino 
acid residues and the resulting electron density is 
readily interpretable. At this stage of refinement 
the root mean square error in the coordinates is 
0.48. The refinement is in progress. 
P-Al-12 
STRUCTURE AND MODE OF A&TiON OF 
ANEWPOTASRiVM~~R 
NORTON RS,I TUDOR =I PALLAGHY PK,* 
PENNlNGTONMW.2 
1 Biomolecular R&arch Institute. Parkville (AUS), 
2 Bachem Bioscience Inc. King of Prussia (USA). 
Purpose: Kv1.3 potassium channels in T- 
lymphocytes are involved in lymphocyte prolifer- 
ation and lvmobokine ruoductiot~ and blockers of 
this char&l &e of i&rest as p&e&l immuno- 
suppressants. ShK toxin (1) is a potent blocker of 
this channel. Our aim is to define the structural 
basis for channel blctckade by thks pblypeptide. 
Methods: The strucmre was solved by 2D NMR. 
Results: ShK toxin shows little sequence similar- 
ity to scorpion-derived potassium channel 
blockers, and its half-cysdnes are paired differ- 
ently. Its structure Pn solution is also different. 
consisting of two shott helices and a series of 
reverse turns. We arf now mapping onto this 
structure the K+ channel binding surface, using 
synthetic analogue da&a, which have identified 
several residues essential for binding (2). Based 
on these data, ShK toxin has also been docked 
into a model of the channel pore (3). 
Conclisfons: ShK toxin constitutes a novel 
protein fold capable of inhibiting potassium 
channel function. 
1. 0. Cast&da et aIt Toafcoa 33,603-613 (1995) 
2. M.W. Pennington et id. Biochem Biophys. Res. 
cononun. 219.6%.701(19%) 
3. J. Aiyaret al. Neuron 13.1169-1181 (1995) 
29 


Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by Heteronuclear NMR Spectroscopy

Dissertations of the University of Groningen

   University of GroningenSolution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined byHeteronuclear NMR SpectroscopyHangyi, Ilona W.; Kroon, Gerard J.A.; Thole, Esther R.; Renken, Remco J.; Dijkstra, Klaas;Scheek, Ruud M.; Robillard, George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:1996Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Hangyi, I. W., Kroon, G. J. A., Thole, E. R., Renken, R. J., Dijkstra, K., Scheek, R. M., & Robillard, G. T.(1996). Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by HeteronuclearNMR Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 11-09-2023P-Al-09 lUNAW.F%AHF@~J.A. KROON, ESTHWRTHOLE,REMCOJ. RENKEN,KLAASDUKS7RA, RUUDM. !$CHEEK and GEORGE T. ROBILLARD. GBB, University of &m&en (NL). l!auym-II mannitol (EIFll) of E coli is part ofa@emthatregulatestheuptakeand phosphorylation of specific carbohydrate subsbks.Ourgroupstudiesthe strutitiction relationship of the cytoplasmic domains of IZ.IF using multidimensional NMRtechniques. The G teminal domain of EV’ (ElIA”“‘, 148 residnes, 16.4 kD) has heen cloned and ovtsqmsd in E cob. The low resoldon strwtwe was determined using 3D NMR techniqnes on ‘%I enriched protein and ‘SN/‘F labeled protein. ELV’ consists of a twwtranded psheet surrounded by 5 cchelices. Mutant ElIAd (ElIAH65Q) that cannot be phosphorylated by phospho-HPr (MIPr) was constructed. The bii interface beti EIIAJkl65Q and I-R is cow to that of EIIAH65Q and P-I-B, using 2D ‘%-HSQC. P-Al-l 1 CRYSTAL STRUCTURE OF THE 13 SUBUNIT HEART CYTOCHROME C OXIDASE. Shinzawa-Itoh, H,I Tsukihara, T,2 Aoyama, H.2 Yamashita, E.2 Tomixaki, T.2 Yamaguchi, H,2 Nakashima. R.1 Yaono, R.1 Yoshikawa , S.1 1Himeji Institute of Technology (JP), 2Institute for Protein Research, Osaka University (JP). Purpose: For elucidation of the reaction mechanism of cytochrome oxidase, crystal structure of the enzyme was solved. Methods: The fully oxidized bovine heart cytochrome c oxidase stabilized with decyl maltoside was crystallized and the crystal structure was solved with MIR method at 2.8 A resolution with an R value of 20.4 % . Results and conclusion: Each structure of all the residues of the monomer (1780 in total) except for 23 residues has been converged to a reasonable structure by structural refinement. A hydrogen bond system including an imidazole bound to CuA. a peptide unit and a propionate of heme a could be an effective electron transfer path between CuA and heme a. Two structures spanned from the cytosolic surfaoe to the matrix including hydrogen bonds and internal cavities likely to contain water molecules, could serve as proton pumping path. Possible channels, for chemical protons to produce H20, for removing the produced H20 and for 02 were identified. Al 3D structures: experimental determination P-Al-10 STR- OF MARE LiWTOF’E~ AT 4.0& RRSOLtiON KAUR P, SH&&A AK, KAR’lWKEYAN S, MITRA SN AND SINGH TP. Depmnt of Biophysics, AlI,lndia Institute of Medical Sciences, New Delhi-l 10 029 (India) Purpose: Lactofenin an iron binding glyco- protein (MW=80,OOODa) has two structural lobes, each housing one Fe’+ and a synergistic COs’- ion. Stucture determination has been carried out to understand the mechanism of action and the functional role played by lactofetin. Method: Purification from mare colostrum/milk by ion exchange and gel filteration. Crystalliza- tion by microdialysis method. structure elucidation using Molecular Replacement. Results:Tbe protein crystallizes in orthorhombic space group ~2~2~2, with a=79.8A, b=lOsSA, ?=112.0A, 2=4 d an a solvent content of 57%. The structure has been refined for 6474 reflec- tions in the resolution range IO-4A. The current R factor is 0.27. The model contains 690 amino acid residues and the resulting electron density is readily interpretable. At this stage of refinement the root mean square error in the coordinates is 0.48. The refinement is in progress. P-Al-12 STRUCTURE AND MODE OF A&TiON OF ANEWPOTASRiVM~~R NORTON RS,I TUDOR =I PALLAGHY PK,* PENNlNGTONMW.2 1 Biomolecular R&arch Institute. Parkville (AUS), 2 Bachem Bioscience Inc. King of Prussia (USA). Purpose: Kv1.3 potassium channels in T- lymphocytes are involved in lymphocyte prolifer- ation and lvmobokine ruoductiot~ and blockers of this char&l &e of i&rest as p&e&l immuno- suppressants. ShK toxin (1) is a potent blocker of this channel. Our aim is to define the structural basis for channel blctckade by thks pblypeptide. Methods: The strucmre was solved by 2D NMR. Results: ShK toxin shows little sequence similar- ity to scorpion-derived potassium channel blockers, and its half-cysdnes are paired differ- ently. Its structure Pn solution is also different. consisting of two shott helices and a series of reverse turns. We arf now mapping onto this structure the K+ channel binding surface, using synthetic analogue da&a, which have identified several residues essential for binding (2). Based on these data, ShK toxin has also been docked into a model of the channel pore (3). Conclisfons: ShK toxin constitutes a novel protein fold capable of inhibiting potassium channel function. 1. 0. Cast&da et aIt Toafcoa 33,603-613 (1995) 2. M.W. Pennington et id. Biochem Biophys. Res. cononun. 219.6%.701(19%) 3. J. Aiyaret al. Neuron 13.1169-1181 (1995) 29 

English

Proceedings - University of Groningen

   University of GroningenSolution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined byHeteronuclear NMR SpectroscopyHangyi, Ilona W.; Kroon, Gerard J.A.; Thole, Esther R.; Renken, Remco J.; Dijkstra, Klaas;Scheek, Ruud M.; Robillard, George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:1996Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Hangyi, I. W., Kroon, G. J. A., Thole, E. R., Renken, R. J., Dijkstra, K., Scheek, R. M., & Robillard, G. T.(1996). Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by HeteronuclearNMR Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 04-06-2022P-Al-09 lUNAW.F%AHF@~J.A. KROON, ESTHWRTHOLE,REMCOJ. RENKEN,KLAASDUKS7RA, RUUDM. !$CHEEK and GEORGE T. ROBILLARD. GBB, University of &m&en (NL). l!auym-II mannitol (EIFll) of E coli is part ofa@emthatregulatestheuptakeand phosphorylation of specific carbohydrate subsbks.Ourgroupstudiesthe strutitiction relationship of the cytoplasmic domains of IZ.IF using multidimensional NMRtechniques. The G teminal domain of EV’ (ElIA”“‘, 148 residnes, 16.4 kD) has heen cloned and ovtsqmsd in E cob. The low resoldon strwtwe was determined using 3D NMR techniqnes on ‘%I enriched protein and ‘SN/‘F labeled protein. ELV’ consists of a twwtranded psheet surrounded by 5 cchelices. Mutant ElIAd (ElIAH65Q) that cannot be phosphorylated by phospho-HPr (MIPr) was constructed. The bii interface beti EIIAJkl65Q and I-R is cow to that of EIIAH65Q and P-I-B, using 2D ‘%-HSQC. P-Al-l 1 CRYSTAL STRUCTURE OF THE 13 SUBUNIT HEART CYTOCHROME C OXIDASE. Shinzawa-Itoh, H,I Tsukihara, T,2 Aoyama, H.2 Yamashita, E.2 Tomixaki, T.2 Yamaguchi, H,2 Nakashima. R.1 Yaono, R.1 Yoshikawa , S.1 1Himeji Institute of Technology (JP), 2Institute for Protein Research, Osaka University (JP). Purpose: For elucidation of the reaction mechanism of cytochrome oxidase, crystal structure of the enzyme was solved. Methods: The fully oxidized bovine heart cytochrome c oxidase stabilized with decyl maltoside was crystallized and the crystal structure was solved with MIR method at 2.8 A resolution with an R value of 20.4 % . Results and conclusion: Each structure of all the residues of the monomer (1780 in total) except for 23 residues has been converged to a reasonable structure by structural refinement. A hydrogen bond system including an imidazole bound to CuA. a peptide unit and a propionate of heme a could be an effective electron transfer path between CuA and heme a. Two structures spanned from the cytosolic surfaoe to the matrix including hydrogen bonds and internal cavities likely to contain water molecules, could serve as proton pumping path. Possible channels, for chemical protons to produce H20, for removing the produced H20 and for 02 were identified. Al 3D structures: experimental determination P-Al-10 STR- OF MARE LiWTOF’E~ AT 4.0& RRSOLtiON KAUR P, SH&&A AK, KAR’lWKEYAN S, MITRA SN AND SINGH TP. Depmnt of Biophysics, AlI,lndia Institute of Medical Sciences, New Delhi-l 10 029 (India) Purpose: Lactofenin an iron binding glyco- protein (MW=80,OOODa) has two structural lobes, each housing one Fe’+ and a synergistic COs’- ion. Stucture determination has been carried out to understand the mechanism of action and the functional role played by lactofetin. Method: Purification from mare colostrum/milk by ion exchange and gel filteration. Crystalliza- tion by microdialysis method. structure elucidation using Molecular Replacement. Results:Tbe protein crystallizes in orthorhombic space group ~2~2~2, with a=79.8A, b=lOsSA, ?=112.0A, 2=4 d an a solvent content of 57%. The structure has been refined for 6474 reflec- tions in the resolution range IO-4A. The current R factor is 0.27. The model contains 690 amino acid residues and the resulting electron density is readily interpretable. At this stage of refinement the root mean square error in the coordinates is 0.48. The refinement is in progress. P-Al-12 STRUCTURE AND MODE OF A&TiON OF ANEWPOTASRiVM~~R NORTON RS,I TUDOR =I PALLAGHY PK,* PENNlNGTONMW.2 1 Biomolecular R&arch Institute. Parkville (AUS), 2 Bachem Bioscience Inc. King of Prussia (USA). Purpose: Kv1.3 potassium channels in T- lymphocytes are involved in lymphocyte prolifer- ation and lvmobokine ruoductiot~ and blockers of this char&l &e of i&rest as p&e&l immuno- suppressants. ShK toxin (1) is a potent blocker of this channel. Our aim is to define the structural basis for channel blctckade by thks pblypeptide. Methods: The strucmre was solved by 2D NMR. Results: ShK toxin shows little sequence similar- ity to scorpion-derived potassium channel blockers, and its half-cysdnes are paired differ- ently. Its structure Pn solution is also different. consisting of two shott helices and a series of reverse turns. We arf now mapping onto this structure the K+ channel binding surface, using synthetic analogue da&a, which have identified several residues essential for binding (2). Based on these data, ShK toxin has also been docked into a model of the channel pore (3). Conclisfons: ShK toxin constitutes a novel protein fold capable of inhibiting potassium channel function. 1. 0. Cast&da et aIt Toafcoa 33,603-613 (1995) 2. M.W. Pennington et id. Biochem Biophys. Res. cononun. 219.6%.701(19%) 3. J. Aiyaret al. Neuron 13.1169-1181 (1995) 29 

NARCIS 

University of Groningen

   University of GroningenSolution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined byHeteronuclear NMR SpectroscopyHangyi, Ilona W.; Kroon, Gerard J.A.; Thole, Esther R.; Renken, Remco J.; Dijkstra, Klaas;Scheek, Ruud M.; Robillard, George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:1996Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Hangyi, I. W., Kroon, G. J. A., Thole, E. R., Renken, R. J., Dijkstra, K., Scheek, R. M., & Robillard, G. T.(1996). Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by HeteronuclearNMR Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 21-05-2020P-Al-09 lUNAW.F%AHF@~J.A. KROON, ESTHWRTHOLE,REMCOJ. RENKEN,KLAASDUKS7RA, RUUDM. !$CHEEK and GEORGE T. ROBILLARD. GBB, University of &m&en (NL). l!auym-II mannitol (EIFll) of E coli is part ofa@emthatregulatestheuptakeand phosphorylation of specific carbohydrate subsbks.Ourgroupstudiesthe strutitiction relationship of the cytoplasmic domains of IZ.IF using multidimensional NMRtechniques. The G teminal domain of EV’ (ElIA”“‘, 148 residnes, 16.4 kD) has heen cloned and ovtsqmsd in E cob. The low resoldon strwtwe was determined using 3D NMR techniqnes on ‘%I enriched protein and ‘SN/‘F labeled protein. ELV’ consists of a twwtranded psheet surrounded by 5 cchelices. Mutant ElIAd (ElIAH65Q) that cannot be phosphorylated by phospho-HPr (MIPr) was constructed. The bii interface beti EIIAJkl65Q and I-R is cow to that of EIIAH65Q and P-I-B, using 2D ‘%-HSQC. P-Al-l 1 CRYSTAL STRUCTURE OF THE 13 SUBUNIT HEART CYTOCHROME C OXIDASE. Shinzawa-Itoh, H,I Tsukihara, T,2 Aoyama, H.2 Yamashita, E.2 Tomixaki, T.2 Yamaguchi, H,2 Nakashima. R.1 Yaono, R.1 Yoshikawa , S.1 1Himeji Institute of Technology (JP), 2Institute for Protein Research, Osaka University (JP). Purpose: For elucidation of the reaction mechanism of cytochrome oxidase, crystal structure of the enzyme was solved. Methods: The fully oxidized bovine heart cytochrome c oxidase stabilized with decyl maltoside was crystallized and the crystal structure was solved with MIR method at 2.8 A resolution with an R value of 20.4 % . Results and conclusion: Each structure of all the residues of the monomer (1780 in total) except for 23 residues has been converged to a reasonable structure by structural refinement. A hydrogen bond system including an imidazole bound to CuA. a peptide unit and a propionate of heme a could be an effective electron transfer path between CuA and heme a. Two structures spanned from the cytosolic surfaoe to the matrix including hydrogen bonds and internal cavities likely to contain water molecules, could serve as proton pumping path. Possible channels, for chemical protons to produce H20, for removing the produced H20 and for 02 were identified. Al 3D structures: experimental determination P-Al-10 STR- OF MARE LiWTOF’E~ AT 4.0& RRSOLtiON KAUR P, SH&&A AK, KAR’lWKEYAN S, MITRA SN AND SINGH TP. Depmnt of Biophysics, AlI,lndia Institute of Medical Sciences, New Delhi-l 10 029 (India) Purpose: Lactofenin an iron binding glyco- protein (MW=80,OOODa) has two structural lobes, each housing one Fe’+ and a synergistic COs’- ion. Stucture determination has been carried out to understand the mechanism of action and the functional role played by lactofetin. Method: Purification from mare colostrum/milk by ion exchange and gel filteration. Crystalliza- tion by microdialysis method. structure elucidation using Molecular Replacement. Results:Tbe protein crystallizes in orthorhombic space group ~2~2~2, with a=79.8A, b=lOsSA, ?=112.0A, 2=4 d an a solvent content of 57%. The structure has been refined for 6474 reflec- tions in the resolution range IO-4A. The current R factor is 0.27. The model contains 690 amino acid residues and the resulting electron density is readily interpretable. At this stage of refinement the root mean square error in the coordinates is 0.48. The refinement is in progress. P-Al-12 STRUCTURE AND MODE OF A&TiON OF ANEWPOTASRiVM~~R NORTON RS,I TUDOR =I PALLAGHY PK,* PENNlNGTONMW.2 1 Biomolecular R&arch Institute. Parkville (AUS), 2 Bachem Bioscience Inc. King of Prussia (USA). Purpose: Kv1.3 potassium channels in T- lymphocytes are involved in lymphocyte prolifer- ation and lvmobokine ruoductiot~ and blockers of this char&l &e of i&rest as p&e&l immuno- suppressants. ShK toxin (1) is a potent blocker of this channel. Our aim is to define the structural basis for channel blctckade by thks pblypeptide. Methods: The strucmre was solved by 2D NMR. Results: ShK toxin shows little sequence similar- ity to scorpion-derived potassium channel blockers, and its half-cysdnes are paired differ- ently. Its structure Pn solution is also different. consisting of two shott helices and a series of reverse turns. We arf now mapping onto this structure the K+ channel binding surface, using synthetic analogue da&a, which have identified several residues essential for binding (2). Based on these data, ShK toxin has also been docked into a model of the channel pore (3). Conclisfons: ShK toxin constitutes a novel protein fold capable of inhibiting potassium channel function. 1. 0. Cast&da et aIt Toafcoa 33,603-613 (1995) 2. M.W. Pennington et id. Biochem Biophys. Res. cononun. 219.6%.701(19%) 3. J. Aiyaret al. Neuron 13.1169-1181 (1995) 29 

University of Groningen Digital Archive

   University of GroningenSolution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined byHeteronuclear NMR SpectroscopyHangyi, Ilona W.; Kroon, Gerard J.A.; Thole, Esther R.; Renken, Remco J.; Dijkstra, Klaas;Scheek, Ruud M.; Robillard, George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:1996Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Hangyi, I. W., Kroon, G. J. A., Thole, E. R., Renken, R. J., Dijkstra, K., Scheek, R. M., & Robillard, G. T.(1996). Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by HeteronuclearNMR Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 05-06-2022P-Al-09 lUNAW.F%AHF@~J.A. KROON, ESTHWRTHOLE,REMCOJ. RENKEN,KLAASDUKS7RA, RUUDM. !$CHEEK and GEORGE T. ROBILLARD. GBB, University of &m&en (NL). l!auym-II mannitol (EIFll) of E coli is part ofa@emthatregulatestheuptakeand phosphorylation of specific carbohydrate subsbks.Ourgroupstudiesthe strutitiction relationship of the cytoplasmic domains of IZ.IF using multidimensional NMRtechniques. The G teminal domain of EV’ (ElIA”“‘, 148 residnes, 16.4 kD) has heen cloned and ovtsqmsd in E cob. The low resoldon strwtwe was determined using 3D NMR techniqnes on ‘%I enriched protein and ‘SN/‘F labeled protein. ELV’ consists of a twwtranded psheet surrounded by 5 cchelices. Mutant ElIAd (ElIAH65Q) that cannot be phosphorylated by phospho-HPr (MIPr) was constructed. The bii interface beti EIIAJkl65Q and I-R is cow to that of EIIAH65Q and P-I-B, using 2D ‘%-HSQC. P-Al-l 1 CRYSTAL STRUCTURE OF THE 13 SUBUNIT HEART CYTOCHROME C OXIDASE. Shinzawa-Itoh, H,I Tsukihara, T,2 Aoyama, H.2 Yamashita, E.2 Tomixaki, T.2 Yamaguchi, H,2 Nakashima. R.1 Yaono, R.1 Yoshikawa , S.1 1Himeji Institute of Technology (JP), 2Institute for Protein Research, Osaka University (JP). Purpose: For elucidation of the reaction mechanism of cytochrome oxidase, crystal structure of the enzyme was solved. Methods: The fully oxidized bovine heart cytochrome c oxidase stabilized with decyl maltoside was crystallized and the crystal structure was solved with MIR method at 2.8 A resolution with an R value of 20.4 % . Results and conclusion: Each structure of all the residues of the monomer (1780 in total) except for 23 residues has been converged to a reasonable structure by structural refinement. A hydrogen bond system including an imidazole bound to CuA. a peptide unit and a propionate of heme a could be an effective electron transfer path between CuA and heme a. Two structures spanned from the cytosolic surfaoe to the matrix including hydrogen bonds and internal cavities likely to contain water molecules, could serve as proton pumping path. Possible channels, for chemical protons to produce H20, for removing the produced H20 and for 02 were identified. Al 3D structures: experimental determination P-Al-10 STR- OF MARE LiWTOF’E~ AT 4.0& RRSOLtiON KAUR P, SH&&A AK, KAR’lWKEYAN S, MITRA SN AND SINGH TP. Depmnt of Biophysics, AlI,lndia Institute of Medical Sciences, New Delhi-l 10 029 (India) Purpose: Lactofenin an iron binding glyco- protein (MW=80,OOODa) has two structural lobes, each housing one Fe’+ and a synergistic COs’- ion. Stucture determination has been carried out to understand the mechanism of action and the functional role played by lactofetin. Method: Purification from mare colostrum/milk by ion exchange and gel filteration. Crystalliza- tion by microdialysis method. structure elucidation using Molecular Replacement. Results:Tbe protein crystallizes in orthorhombic space group ~2~2~2, with a=79.8A, b=lOsSA, ?=112.0A, 2=4 d an a solvent content of 57%. The structure has been refined for 6474 reflec- tions in the resolution range IO-4A. The current R factor is 0.27. The model contains 690 amino acid residues and the resulting electron density is readily interpretable. At this stage of refinement the root mean square error in the coordinates is 0.48. The refinement is in progress. P-Al-12 STRUCTURE AND MODE OF A&TiON OF ANEWPOTASRiVM~~R NORTON RS,I TUDOR =I PALLAGHY PK,* PENNlNGTONMW.2 1 Biomolecular R&arch Institute. Parkville (AUS), 2 Bachem Bioscience Inc. King of Prussia (USA). Purpose: Kv1.3 potassium channels in T- lymphocytes are involved in lymphocyte prolifer- ation and lvmobokine ruoductiot~ and blockers of this char&l &e of i&rest as p&e&l immuno- suppressants. ShK toxin (1) is a potent blocker of this channel. Our aim is to define the structural basis for channel blctckade by thks pblypeptide. Methods: The strucmre was solved by 2D NMR. Results: ShK toxin shows little sequence similar- ity to scorpion-derived potassium channel blockers, and its half-cysdnes are paired differ- ently. Its structure Pn solution is also different. consisting of two shott helices and a series of reverse turns. We arf now mapping onto this structure the K+ channel binding surface, using synthetic analogue da&a, which have identified several residues essential for binding (2). Based on these data, ShK toxin has also been docked into a model of the channel pore (3). Conclisfons: ShK toxin constitutes a novel protein fold capable of inhibiting potassium channel function. 1. 0. Cast&da et aIt Toafcoa 33,603-613 (1995) 2. M.W. Pennington et id. Biochem Biophys. Res. cononun. 219.6%.701(19%) 3. J. Aiyaret al. Neuron 13.1169-1181 (1995) 29 

ARTS repository - University of Groningen

   University of GroningenSolution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined byHeteronuclear NMR SpectroscopyHangyi, Ilona W.; Kroon, Gerard J.A.; Thole, Esther R.; Renken, Remco J.; Dijkstra, Klaas;Scheek, Ruud M.; Robillard, George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:1996Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Hangyi, I. W., Kroon, G. J. A., Thole, E. R., Renken, R. J., Dijkstra, K., Scheek, R. M., & Robillard, G. T.(1996). Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by HeteronuclearNMR Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 01-11-2023P-Al-09 lUNAW.F%AHF@~J.A. KROON, ESTHWRTHOLE,REMCOJ. RENKEN,KLAASDUKS7RA, RUUDM. !$CHEEK and GEORGE T. ROBILLARD. GBB, University of &m&en (NL). l!auym-II mannitol (EIFll) of E coli is part ofa@emthatregulatestheuptakeand phosphorylation of specific carbohydrate subsbks.Ourgroupstudiesthe strutitiction relationship of the cytoplasmic domains of IZ.IF using multidimensional NMRtechniques. The G teminal domain of EV’ (ElIA”“‘, 148 residnes, 16.4 kD) has heen cloned and ovtsqmsd in E cob. The low resoldon strwtwe was determined using 3D NMR techniqnes on ‘%I enriched protein and ‘SN/‘F labeled protein. ELV’ consists of a twwtranded psheet surrounded by 5 cchelices. Mutant ElIAd (ElIAH65Q) that cannot be phosphorylated by phospho-HPr (MIPr) was constructed. The bii interface beti EIIAJkl65Q and I-R is cow to that of EIIAH65Q and P-I-B, using 2D ‘%-HSQC. P-Al-l 1 CRYSTAL STRUCTURE OF THE 13 SUBUNIT HEART CYTOCHROME C OXIDASE. Shinzawa-Itoh, H,I Tsukihara, T,2 Aoyama, H.2 Yamashita, E.2 Tomixaki, T.2 Yamaguchi, H,2 Nakashima. R.1 Yaono, R.1 Yoshikawa , S.1 1Himeji Institute of Technology (JP), 2Institute for Protein Research, Osaka University (JP). Purpose: For elucidation of the reaction mechanism of cytochrome oxidase, crystal structure of the enzyme was solved. Methods: The fully oxidized bovine heart cytochrome c oxidase stabilized with decyl maltoside was crystallized and the crystal structure was solved with MIR method at 2.8 A resolution with an R value of 20.4 % . Results and conclusion: Each structure of all the residues of the monomer (1780 in total) except for 23 residues has been converged to a reasonable structure by structural refinement. A hydrogen bond system including an imidazole bound to CuA. a peptide unit and a propionate of heme a could be an effective electron transfer path between CuA and heme a. Two structures spanned from the cytosolic surfaoe to the matrix including hydrogen bonds and internal cavities likely to contain water molecules, could serve as proton pumping path. Possible channels, for chemical protons to produce H20, for removing the produced H20 and for 02 were identified. Al 3D structures: experimental determination P-Al-10 STR- OF MARE LiWTOF’E~ AT 4.0& RRSOLtiON KAUR P, SH&&A AK, KAR’lWKEYAN S, MITRA SN AND SINGH TP. Depmnt of Biophysics, AlI,lndia Institute of Medical Sciences, New Delhi-l 10 029 (India) Purpose: Lactofenin an iron binding glyco- protein (MW=80,OOODa) has two structural lobes, each housing one Fe’+ and a synergistic COs’- ion. Stucture determination has been carried out to understand the mechanism of action and the functional role played by lactofetin. Method: Purification from mare colostrum/milk by ion exchange and gel filteration. Crystalliza- tion by microdialysis method. structure elucidation using Molecular Replacement. Results:Tbe protein crystallizes in orthorhombic space group ~2~2~2, with a=79.8A, b=lOsSA, ?=112.0A, 2=4 d an a solvent content of 57%. The structure has been refined for 6474 reflec- tions in the resolution range IO-4A. The current R factor is 0.27. The model contains 690 amino acid residues and the resulting electron density is readily interpretable. At this stage of refinement the root mean square error in the coordinates is 0.48. The refinement is in progress. P-Al-12 STRUCTURE AND MODE OF A&TiON OF ANEWPOTASRiVM~~R NORTON RS,I TUDOR =I PALLAGHY PK,* PENNlNGTONMW.2 1 Biomolecular R&arch Institute. Parkville (AUS), 2 Bachem Bioscience Inc. King of Prussia (USA). Purpose: Kv1.3 potassium channels in T- lymphocytes are involved in lymphocyte prolifer- ation and lvmobokine ruoductiot~ and blockers of this char&l &e of i&rest as p&e&l immuno- suppressants. ShK toxin (1) is a potent blocker of this channel. Our aim is to define the structural basis for channel blctckade by thks pblypeptide. Methods: The strucmre was solved by 2D NMR. Results: ShK toxin shows little sequence similar- ity to scorpion-derived potassium channel blockers, and its half-cysdnes are paired differ- ently. Its structure Pn solution is also different. consisting of two shott helices and a series of reverse turns. We arf now mapping onto this structure the K+ channel binding surface, using synthetic analogue da&a, which have identified several residues essential for binding (2). Based on these data, ShK toxin has also been docked into a model of the channel pore (3). Conclisfons: ShK toxin constitutes a novel protein fold capable of inhibiting potassium channel function. 1. 0. Cast&da et aIt Toafcoa 33,603-613 (1995) 2. M.W. Pennington et id. Biochem Biophys. Res. cononun. 219.6%.701(19%) 3. J. Aiyaret al. Neuron 13.1169-1181 (1995) 29 

Solution Structure of EIIAmtl and Interactions of EIIAmtl with HPr Determined by Heteronuclear NMR Spectroscopy

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