Title

Expression, purification, and characterization of thymidylate synthase from Lactococcus lactis

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Publication Date

1994-07-01

Document Type

Article

Subjects

Amino Acid Sequence; Binding Sites; Crystallization; Escherichia coli; *Gene Expression; Hydrogen-Ion Concentration; Lactococcus lactis; Models, Molecular; Molecular Sequence Data; Molecular Structure; Osmolar Concentration; Recombinant Proteins; Structure-Activity Relationship; Thymidylate Synthase

Disciplines

Biochemistry, Biophysics, and Structural Biology | Pharmacology, Toxicology and Environmental Health

Abstract

The thymidylate synthase (TS) gene from Lactococcus lactis has been highly expressed in Escherichia coli. The TS protein was purified by sequential chromatography on Q-Sepharose and phenyl-Sepharose. Six grams of cell pellet yielded 140 mg of homogeneous TS. TS is a highly conserved enzyme, and several of the conserved amino acid residues that have been implicated in catalytic function are altered in L. lactis TS. By use of a 3-dimensional homology model, we have predicted covariant changes that might compensate for these differences. With the large amounts of L. lactis TS now available, studies can be pursued to understand the structure-function relationships of this enzyme compared to other TSs and to confirm the presumed roles of the compensatory changes predicted in the homology model.

DOI of Published Version

10.1002/pro.5560030715

Source

Protein Sci. 1994 Jul;3(7):1114-6. Link to article on publisher's site

Journal/Book/Conference Title

Protein science : a publication of the Protein Society

Related Resources

Link to Article in PubMed

PubMed ID

7920258

Link to Full Text

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