A thermostable K+‐stimulated vacuolar‐type pyrophosphatase from the hyperthermophilic bacterium Thermotoga maritima

FEBS Letters

Losada, Manuel¹  López-Marqués, Rosa L.¹  Pérez-Castiñeira, José R.¹  Serrano, Aurelio¹ 

[1] Instituto de Bioquı́mica Vegetal y Fotosı́ntesis (Universidad de Sevilla-CSIC), Avda Americo Vespucio s/n, 41092 Sevilla, Spain

关键词: Heterologous expression;    K+-stimulation;    Vacuolar-type H+-pyrophosphatase;    Saccharomyces cerevisiae;    Thermotoga maritima;    PPi;    inorganic pyrophosphate;    PCR;    polymerase chain reaction;    RVP;    Rhodospirillum rubrum V-PPase;    TVP;    Thermotoga maritima V-PPase;    V-PPase;    vacuolar-type inorganic pyrophosphatase;   

DOI  :  10.1016/S0014-5793(01)02390-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Current evidence suggests the occurrence of two classes of vacuolar-type H⁺-translocating inorganic pyrophosphatases (V-PPases): K⁺-insensitive proteins, identified in eukaryotes, bacteria and archaea, and K⁺-stimulated V-PPases, identified to date only in eukaryotes. Here, we describe the functional characterization of a thermostable V-PPase from the anaerobic hyperthermophilic bacterium Thermotoga maritima by heterologous expression in Saccharomyces cerevisiae. The activity of this 71-kDa membrane-embedded polypeptide has a near obligate requirement for K⁺, like the plant V-PPase, and its thermostability depends on the binding of Mg²⁺. Phylogenetic analysis of protein sequences consistently assigned the T. maritima V-PPase to the K⁺-sensitive class of V-PPases so far only known for eukaryotes. The finding of a K⁺-stimulated V-PPase also in a member of a primitive eubacterial lineage strongly supports an ancient evolutionary origin of this group of pyrophosphate-energized proton pumps.

【 授权许可】

Unknown