Analysis of the regulatory domain of yeast plasma membrane H+‐ATPase by directed mutagenesis and intragenic suppression

FEBS Letters

Serrano, Ramon²  Eraso, Pilar¹  Portillo, Francisco¹ 

[1] Instituto de Investigaciones Biomedicas, Departamento de Bioquimica, Facultad de Medicina, Universidad Autonoma, Arluro Duperier 4, 28029 Madrid, Spain;European Molecular Biology Laboratory, Meyerhofstrasse 1, 6900 Heidelberg, Germany

关键词: H+-ATPase;    Plasma membrane;    Domain interaction;    Site-directed mutagenesis;    Saccharomyces cerevisiae;   

DOI  :  10.1016/0014-5793(91)80018-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The yeast plasma membrane H⁺-ATPase is activated in vivo by glucose metabolism, and previous deletion analysis has shown the C-terminus of the enzyme to be involved in this regulation. Site-directed mutagenesis demonstrates that Arg⁹⁰⁹ and Thr⁹¹² at the C-terminus are important for the increase in V _max of the ATPase induced by glucose. Other changes in kinetic parameters induced by glucose are largely independent of these ammo acids. Arg⁹⁰⁹ and Thr⁹¹² form a potential phosphorylation site for calmodulin-dependent multiprotein kinase. A double mutation of Ser⁹¹¹ and Thr⁹¹² to Ala results in no cell growth in glucose medium and greatly reduced activation of the ATPase by glucose. Growth and activity are restored by a third mutation (Ala⁵⁴⁷ → Val) at the catalytic domain, providing genetic evidence for domain interaction.

【 授权许可】

Unknown