Conformational gel analysis and graphics: Measurement of side chain rotational isomer populations by NMR and molecular mechanics

Physics – Biological Physics

Scientific paper

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9 pages, 6 figures, REVTeX v4

Scientific paper

Conformational gel analysis and graphics systematically identifies and evaluates plausible alternatives to the side chain conformations found by conventional peptide or protein structure determination methods. The proposed analysis determines the populations of side chain rotational isomers and the probability distribution of these populations. The following steps are repeated for each side chain of a peptide or protein: first, extract the local molecular mechanics of side chain rotational isomerization from a single representative global conformation; second, expand the predominant set of rotational isomers to include all probable rotational isomers down to those that constitute just a small percentage of the population; and third, evaluate the constraints vicinal coupling constants and NOESY cross relaxation rates place on rotational isomer populations. In this article we apply conformational gel analysis to the cobalt glycyl-leucine dipeptide and detail the steps necessary to generalize the analysis to other amino acid side chains in other peptides and proteins. For a side chain buried within a protein interior, it is noteworthy that the set of probable rotational isomers may contain one or more rotational isomers that are not identified by conventional NMR structure determination methods. In cases such as this the conformational gel graphics fully accounts for the interplay of molecular mechanics and NMR data constraints on the population estimates. The analysis is particularly suited to identifying side chain rotational isomers that constitute a small percentage of the population, but nevertheless might be structurally and functionally very significant.

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