Active-site-mutagenesis Study of Rat Liver Betaine-homocysteine S-methyltransferase

Overview

Journal Biochem J

Specialty Biochemistry

Date 2002 Dec 19

PMID 12487625

Citations 9

Authors

Beatriz Gonzalez

Nuria Campillo

Francisco Garrido

Maria Gasset

Juliana Sanz-Aparicio

Maria A Pajares

Affiliations

Soon will be listed here.

Abstract

A site-directed-mutagenesis study of putative active-site residues in rat liver betaine-homocysteine S-methyltransferase has been carried out. Identification of these amino acids was based on data derived from a structural model of the enzyme. No alterations in the CD spectra or the gel-filtration chromatography elution pattern were observed with the mutants, thus suggesting no modification in the secondary structure content or in the association state of the proteins. All the mutants obtained showed a reduction of the enzyme activity, the most dramatic effect being that of Glu(159), followed by Tyr(77) and Asp(26). Changes in affinity for either of the substrates, homocysteine or betaine, were detected when substitutions were performed of Glu(21), Asp(26), Phe(74) and Cys(186). Interestingly, Asp(26), postulated to be involved in homocysteine binding, has a strong effect on affinity for betaine. The relevance of these results is discussed in the light of very recent structural data obtained for the human enzyme.

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