13 research outputs found
Automated concept and relationship extraction for the semi-automated ontology management (SEAM) system
An effective potential for calculating free energies .1. General concepts and approximations.
Analysis of three human interleukin 5 structures suggests a possible receptor binding mechanism.
AbstractWe compared three crystal structures of human interleukin 5 (hIL5) expressed in either E. coli (hIL5E.coli), Sf9 cells (hIL5Sf9) or Drosophila cells (hIL5Drosophila). The dimeric hIL5 structures show subtle but significant conformational differences which are probably a consequence of the different crystallization conditions trapping this protein into one of two states. We refer to these two distinct conformations as the `open' and `tight' state, according to the packing around the cleft between the two subunits. We hypothesize that these two stable conformational states reflect the structure of the free or receptor bound hIL5
Structural analysis and identification of gel-purified proteins, available in the femtomole range, using a novel computer program for peptide sequence assignment, by matrix-assisted laser desorption ionization reflectron time-of-flight mass spectrometry.
A proposed architecture for lecithin cholesterol acyl transferase (LCAT): Identification of the catalytic triad and molecular modeling.
Characterization of functional residues in the interfacial recognition domain of lecithin cholesterol acyltransferase (LCAT).
Characterization Of Functional Residues In The Interfacial Recognition Domain Of Lecithin Cholesterol Acyltransferase (Lcat)
Lecithin cholesterol acyltransferase (LCAT) is an interfacial enzyme active on
both high-density (HDL) and low-density lipoproteins (LDL). Threading alignments
of LCAT with lipases suggest that residues 50-74 form an interfacial recognition
site and this hypothesis was tested by site-directed mutagenesis. The
(delta56-68) deletion mutant had no activity on any substrate. Substitution of
W61 with F, Y, L or G suggested that an aromatic residue is required for full
enzymatic activity. The activity of the W61F and W61Y mutants was retained on HDL
but decreased on LDL, possibly owing to impaired accessibility to the LDL lipid
substrate. The decreased activity of the single R52A and K53A mutants on HDL and
LDL and the severer effect of the double mutation suggested that these conserved
residues contribute to the folding of the LCAT lid. The membrane-destabilizing
properties of the LCAT 56-68 helical segment were demonstrated using the
corresponding synthetic peptide. An M65N-N66M substitution decreased both the
fusogenic properties of the peptide and the activity of the mutant enzyme on all
substrates. These results suggest that the putative interfacial recognition
domain of LCAT plays an important role in regulating the interaction of the
enzyme with its organized lipoprotein substrates