Crystallization and preliminary X-ray analysis of a bacterial psychrophilic enzyme, phosphoglycerate kinase.
The glycolytic enzyme phosphoglycerate kinase (PGK) from the Antarctic microorganism Pseudomonas sp. TACII18 is a cold-adapted enzyme that displays a high specific activity at low temperatures and decreased thermostability relative to its mesophilic counterpart. Herein, the preliminary crystallization and structure solution of psychrophilic PGK in its native form and cocrystallized with 3-phosphoglyceric acid (3-PGA) and the ATP analogue adenylyl imidophosphate (AMP-PNP) is reported. The complexed form of PGK crystallized in 2-3 d at 290 K, whereas the native form of the enzyme required 8-12 months. Morphologically, both crystal forms are similar and X-ray diffraction experiments indicate that the crystals are isomorphous. The crystals diffracted to a resolution of 2.0 A and belong to the space group P3(2). with unit-cell parameters a = b = 58.5, c = 85.4 A.
Mandelman
D
D
Institut de Biologie et Chimie des Protéines, UMR 5086 Laboratoire de Bio-cristallographie, CNRS - Université Claude Bernard Lyon I, 69367 Lyon CEDEX 07, France.
Bentahir
M
M
Feller
G
G
Gerday
C
C
Haser
R
R
D003460:Crystallization; D018360:Crystallography, X-Ray; D010735:Phosphoglycerate Kinase / Q000737:chemistry*; D011487:Protein Conformation; D011549:Pseudomonas / Q000201:enzymology*
D010735:Phosphoglycerate Kinase