Open AccessBioenergetic Analysis of FLAG Tagged-Subunit 8 of Saccharomyces cerevisiae Mitochondrial ATP Synthase
Author(s) -
I Made Artika
Publication year - 2010
Publication title -
microbiology indonesia
Language(s) - English
Resource type - Journals
eISSN - 2087-8575
pISSN - 1978-3477
DOI - 10.5454/mi.4.3.2
Subject(s) - atp synthase , atp synthase gamma subunit , protein subunit , bioenergetics , saccharomyces cerevisiae , mitochondrion , biology , adenosine triphosphate , biochemistry , v atpase , yeast , inner mitochondrial membrane , atpase , gene , enzyme , atp hydrolysis
The majority of cellular energy in the form of adenosine triphosphate (ATP) is synthesized by the F1F0-ATP synthase. The yeast mitochondrial F1F0-ATP synthase is a multisubunit complex that contains at least 17 different subunits grouped into F1 and F0 sectors. Subunit 8 of yeast mitochondrial ATP synthase is a hydrophobic protein of 48 amino acids encoded by the mitochondrial ATP8 gene. Subunit 8 has three distinct domains; an N-terminal domain, a central hydrophobic domain and a C-terminal domain. FLAG tag addition to the C-terminus of subunit 8 and its variants has facilitated elucidation of subunit 8's membrane topology. In order to analyze its detailed structure and function, a set of strains expressing FLAG tagged-subunit 8 and its variants were subjected to bioenergetic analysis at cellular and mitochondrial levels. Results obtained showed that the hydrophobic character of the central hydrophobic domain of subunit 8 is essential for functional coupling between F1 and F0 sectors, hence for mitochondrial ATP synthase function