179 research outputs found
An Investigation of the Catalytic Mechanism of S-adenosylmethionine Synthetase by QM/MM Calculations
Catalysis by S-adenosylmethionine synthetase has been investigated by quantum mechanical/molecular mechanical calculations, exploiting structures of the active crystalline enzyme. The transition state energy of +19.1 kcal/mol computed for a nucleophilic attack of the methionyl sulfur on carbon-5′ of the nucleotide was indistinguishable from the experimental (solution) value when the QM residues were an uncharged histidine that hydrogen bonds to the leaving oxygen-5′ and an aspartate that chelates a Mg2+ ion, and was similar (+18.8 kcal/mol) when the QM region also included the active site arginine and lysines. The computed energy difference between reactant and product was also consistent with their equimolar abundance in co-crystals. The calculated geometrical changes support catalysis of a SN2 reaction through hydrogen bonding of the liberated oxygen-5′ to the histidine, charge neutralization by the 2 Mg2+ ions, and stabilization of the product sulfonium cation through a close, non-bonded, contact between the sulfur and the ribose 4′-oxygen
Engineering of Cyclodextrin Product Specificity and pH Optima of the Thermostable Cyclodextrin Glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1
The product specificity and pH optimum of the thermostable cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacterium thermosulfurigenes EM1 was engineered using a combination of x-ray crystallography and site-directed mutagenesis. Previously, a crystal soaking experiment with the Bacillus circulans strain 251 β-CGTase had revealed a maltononaose inhibitor bound to the enzyme in an extended conformation. An identical experiment with the CGTase from T. thermosulfurigenes EM1 resulted in a 2.6-Å resolution x-ray structure of a complex with a maltohexaose inhibitor, bound in a different conformation. We hypothesize that the new maltohexaose conformation is related to the enhanced α-cyclodextrin production of the CGTase.
The detailed structural information subsequently allowed engineering of the cyclodextrin product specificity of the CGTase from T. thermosulfurigenes EM1 by site-directed mutagenesis. Mutation D371R was aimed at hindering the maltohexaose conformation and resulted in enhanced production of larger size cyclodextrins (β- and γ-CD). Mutation D197H was aimed at stabilization of the new maltohexaose conformation and resulted in increased production of α-CD.
Glu258 is involved in catalysis in CGTases as well as α-amylases, and is the proton donor in the first step of the cyclization reaction. Amino acids close to Glu258 in the CGTase from T. thermosulfurigenes EM1 were changed. Phe284 was replaced by Lys and Asn327 by Asp. The mutants showed changes in both the high and low pH slopes of the optimum curve for cyclization and hydrolysis when compared with the wild-type enzyme. This suggests that the pH optimum curve of CGTase is determined only by residue Glu258.
Force-Extension Relations for Polymers with Sliding Links
Topological entanglements in polymers are mimicked by sliding rings
(slip-links) which enforce pair contacts between monomers. We study the
force-extension curve for linear polymers in which slip-links create additional
loops of variable size. For a single loop in a phantom chain, we obtain exact
expressions for the average end-to-end separation: The linear response to a
small force is related to the properties of the unstressed chain, while for a
large force the polymer backbone can be treated as a sequence of Pincus--de
Gennes blobs, the constraint effecting only a single blob. Generalizing this
picture, scaling arguments are used to include self-avoiding effects.Comment: 4 pages, 5 figures; accepted to Phys. Rev. E (Brief Report
Abundance of unknots in various models of polymer loops
A veritable zoo of different knots is seen in the ensemble of looped polymer
chains, whether created computationally or observed in vitro. At short loop
lengths, the spectrum of knots is dominated by the trivial knot (unknot). The
fractional abundance of this topological state in the ensemble of all
conformations of the loop of segments follows a decaying exponential form,
, where marks the crossover from a mostly unknotted
(ie topologically simple) to a mostly knotted (ie topologically complex)
ensemble. In the present work we use computational simulation to look closer
into the variation of for a variety of polymer models. Among models
examined, is smallest (about 240) for the model with all segments of the
same length, it is somewhat larger (305) for Gaussian distributed segments, and
can be very large (up to many thousands) when the segment length distribution
has a fat power law tail.Comment: 13 pages, 6 color figure
Tightness of slip-linked polymer chains
We study the interplay between entropy and topological constraints for a
polymer chain in which sliding rings (slip-links) enforce pair contacts between
monomers. These slip-links divide a closed ring polymer into a number of
sub-loops which can exchange length between each other. In the ideal chain
limit, we find the joint probability density function for the sizes of segments
within such a slip-linked polymer chain (paraknot). A particular segment is
tight (small in size) or loose (of the order of the overall size of the
paraknot) depending on both the number of slip-links it incorporates and its
competition with other segments. When self-avoiding interactions are included,
scaling arguments can be used to predict the statistics of segment sizes for
certain paraknot configurations.Comment: 10 pages, 6 figures, REVTeX
Two-stage directed self-assembly of a cyclic [3]catenane.
Interlocked molecules possess properties and functions that depend upon their intricate connectivity. In addition to the topologically trivial rotaxanes, whose structures may be captured by a planar graph, the topologically non-trivial knots and catenanes represent some of chemistry's most challenging synthetic targets because of the three-dimensional assembly necessary for their construction. Here we report the synthesis of a cyclic [3]catenane, which consists of three mutually interpenetrating rings, via an unusual synthetic route. Five distinct building blocks self-assemble into a heteroleptic triangular framework composed of two joined Fe(II)3L3 circular helicates. Subcomponent exchange then enables specific points in the framework to be linked together to generate the cyclic [3]catenane product. Our method represents an advance both in the intricacy of the metal-templated self-assembly procedure and in the use of selective imine exchange to generate a topologically complex product.This work was supported by the UK Engineering and Physical Sciences Research Council (EPSRC) and a Marie Curie fellowship for J.J.H. (ITN-2010–264645). The authors thank the Diamond Light Source (UK) for synchrotron beamtime on I19 (MT7984 and MT8464).This is the author accepted manuscript. The final version is available from NPG via http://dx.doi.org/10.1038/nchem.220
M3B: A Coarse Grain Force Field for Molecular Simulations of Malto-Oligosaccharides and Their Water Mixtures
Thermochemical and Theoretical Studies of Dimethylpyridine-2,6-dicarboxylate and Pyridine-2,3-, Pyridine-2,5-, and Pyridine-2,6-dicarboxylic Acids
Stereochemical Effects in Supramolecular Self-Assembly at Surfaces: 1-D versus 2-D Enantiomorphic Ordering for PVBA and PEBA on Ag(111)
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