Crystallization of acetate kinase from Methanosarcina thermophila and prediction of its fold

✍ Scribed by Kathryn A. Buss; David A. Sanders; Miriam S. Hasson; Cheryl Ingram-Smith; James G. Ferry

• Book ID: 105356500
• Publisher: Cold Spring Harbor Laboratory Press
• Year: 2008
• Tongue: English
• Weight: 375 KB
• Volume: 6
• Category: Article
• ISSN: 0961-8368
• DOI: 10.1002/pro.5560061222

✦ Synopsis

Abstract

The unique biochemical properties of acetate kinase present a classic conundrum in the study of the mechanism of enzyme‐catalyzed phosphoryl transfer. Large, single crystals of acetate kinase from Methanosarcina thermophila were grown from a solution of ammonium sulfate in the presence of ATP. The crystals diffract to beyond 1.7 Å resolution. Analysis of X‐ray data from the crystals is consistent with a space group of __C__2 and unit cell dimensions a = 181 Å, b = 67 Å, c = 83 Å, β = 103°. Diffraction data have been collected from the crystals at 110 and 277 K. Data collected at 277 K extend to lower resolution, but are more reproducible. The orientation of a noncrystallographic twofold axis of symmetry has been determined. Based on an analysis of the predicted amino acid sequences of acetate kinase from several organisms, we hypothesize that acetate kinase is a member of the sugar kinase/actin/hsp70 structural family.