Journal of Biological Chemistry
Volume 273, Issue 4, 23 January 1998, Pages 2199-2206
Journal home page for Journal of Biological Chemistry

PROTEIN CHEMISTRY AND STRUCTURE
Triose-phosphate Isomerase (TIM) of the Psychrophilic BacteriumVibrio marinus: KINETIC AND STRUCTURAL PROPERTIES*

https://doi.org/10.1074/jbc.273.4.2199Get rights and content
Under a Creative Commons license
open access

The purification and characterization of triose-phosphate isomerase from the psychrophilic bacteriumVibrio marinus (vTIM) is described. Crystal structures of the vTIM-sulfate complex and the vTIM-2-phosphoglycolate complex (at a 2.7-Å resolution) are also presented. The optimal growth temperature of Vibrio marinus is 15 °C. Stability studies show that vTIM is an unstable protein with a half-life of only 10 min at 25 °C. The vTIM sequence is most closely related to the sequence ofEscherichia coli TIM (eTIM) (66% identity), and several unique structural features described for eTIM are also seen in vTIM, but eTIM is considerably more stable. The T d values of vTIM and eTIM, determined by calorimetric studies, are 41 and 54 °C, respectively. Amino acid sequence comparison reveals that vTIM has an alanine in loop 8 (at position 238), whereas all other TIM sequences known to date have a serine. The vTIM mutant A238S was produced and characterized. Compared with wild type, the catalytic efficiency of the A238S mutant is somewhat reduced, and its stability is considerably increased.

Cited by (0)

A research associate of the Belgian National Science Foundation (FWO)

*

This work was supported by Services Fédéraux des Affaires Scientifiques, Techniques, et Culturelles Grant PAI P3-044, European Union Grants BIOT CT90-0182 and BIO4-CT96-0670, and Belgian National Science Foundation Grant FRFC 2.4545.96.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) U48935.

The atomic coordinates and structure factors (1AW1 and 1AW2) have been deposited in the Protein Data Bank, Brookhaven National Laboratory, Upton, NY.