Loop movement and catalysis in creatine kinase.

Abstract:

:Recently the crystal structure of creatine kinase from Torpedocalifornica was determined to 2.1 A. The dimeric structure revealed two different forms in the unit cell: one monomer was bound to a substrate, MgADP, and the other monomer was bound to a transition-state analogue complex composed of MgADP, nitrate and creatine. The most striking difference between the structures is the movement of two loops (comprising residues 60-70 and residues 323-333) into the active site in the transition state structure. This loop movement effectively occludes the active site from solvent, and the loops appear to be locked into place by a salt bridge formed between His66 and Asp326. His66 is of particular interest as it is located within a PGHP motif conserved in all creatine kinases but not found in other guanidino kinases. We have carried out alanine-scanning mutagenesis of each of the residues in the PGHP motif and determined that only the His66 plays a significant role in the creatine kinase reaction. Although neither residue interacts directly with the substrate, the interaction His66 and Asp326 appears to be important in providing the precise alignment of substrates necessary for phosphoryl group transfer. Finally, it is clear that neither His66 nor Asp326 are responsible for the pKs observed in the pH-rate profile for HMCK.

journal_name

IUBMB Life

journal_title

IUBMB life

authors

Wang PF,Flynn AJ,McLeish MJ,Kenyon GL

doi

10.1080/15216540500091999

keywords:

subject

Has Abstract

pub_date

2005-04-01 00:00:00

pages

355-62

issue

4-5

eissn

1521-6543

issn

1521-6551

pii

U598150747662283

journal_volume

57

pub_type

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