His(73), often methylated, is an important structural determinant for actin. A mutagenic analysis of HIS(73) of yeast actin

Xiaoyi Yao; Stephanie  Grade; Willy Wriggers; Peter A Rubenstein

doi:10.1074/jbc.274.52.37443

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His(73), often methylated, is an important structural determinant for actin. A mutagenic analysis of HIS(73) of yeast actin

Journal article

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His(73), often methylated, is an important structural determinant for actin. A mutagenic analysis of HIS(73) of yeast actin

Xiaoyi Yao, Stephanie Grade, Willy Wriggers and Peter A Rubenstein

The Journal of biological chemistry, Vol.274(52), pp.37443-37449

12/24/1999

DOI: 10.1074/jbc.274.52.37443

PMID: 10601317

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Abstract

His(73), has been proposed to regulate the release of P(i) from the interior of actin following polymerization-dependent hydrolysis of bound ATP. Although it is a 3-methylhistidine in the vast majority of actins, His(73) is unmethylated in S. cerevisiae actin. We mutated His(73) in yeast actin to Arg, Lys, Ala, Gln, and Glu and detected no altered phenotypes associated with the mutations in vivo. However, they significantly affect actin function in vitro. Substitution of the more basic residues resulted in enhanced thermal stability, decreased rate of nucleotide exchange, and decreased susceptibility to controlled proteolysis relative to wild-type actin. The opposite effects are observed with the neutral and anionic substitutions. All mutations reduced the rate of polymerization. Molecular dynamics simulations predict a new conformation for the His(73) imidazole in the absence of a methyl group. It also predicts that Arg(73) tightens and stabilizes the actin and that Glu(73) causes a rearrangement of the bottom of actin's interdomain cleft leading possibly to our observed destabilization of actin. Considering the exterior location of His(73), this work indicates a surprisingly important role for the residue as a major structural determinant of actin and provides a clue to the impact caused by methylation of His(73).

Temperature

Histidine

Mutagenesis, Site-Directed

Actins - chemistry

Models, Molecular

Polymers - chemistry

Protein Conformation

Methylation

Saccharomyces cerevisiae - chemistry

Details

Title: Subtitle: His(73), often methylated, is an important structural determinant for actin. A mutagenic analysis of HIS(73) of yeast actin
Creators: Xiaoyi Yao - Department of Biochemistry, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA
Stephanie Grade
Willy Wriggers
Peter A Rubenstein
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.274(52), pp.37443-37449
DOI: 10.1074/jbc.274.52.37443
PMID: 10601317
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: GM33689 / NIGMS NIH HHS
Language: English
Date published: 12/24/1999
Academic Unit: Stead Family Department of Pediatrics; Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9984024410702771

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