Open AccessF 1 ‐ATPase from Micrococcus sp. ATCC 398
Author(s) -
RISI Sergio,
HÖCKEL Michael,
HULLA Franz W.,
DOSE Klaus
Publication year - 1977
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1977.tb11931.x
Subject(s) - proteolysis , chemistry , ionic strength , atpase , electrophoresis , sepharose , enzyme , chromatography , gel electrophoresis , protein subunit , molecular mass , polyacrylamide gel electrophoresis , biochemistry , organic chemistry , gene , aqueous solution
The preparation of highly purified F 1 ‐ATPase from Micrococcus sp. ATCC 398 by application of DEAE‐Sepharose CL‐6B chromatography as final step is described. This enzyme consists of five subunits of different molecular weight: α (65000), β (55000), γ (35000), δ (20000), and ∈ (17000). Disc electrophoresis on 5% polyacrylamide gels removes the ∈‐polypeptide yielding an active ATPase complex with four different subunits: α, β, γ, δ. Additionally, by variation of the ionic strength δ can be (partly) removed allowing the isolation by disc electrophoresis of an active ATPase complex which consists only of three different subunits α, β, and γ. If the DEAE‐Sepharose chromatography is carried out in the absence of diisopropyl phosphofluoridate (auto)proteolysis yields both an active ATPase with the subunits α + (mol. wt 61000), β, γ, and δ and an inactive protein complex with the subunits α + , β, γ, δ, and two additional polypeptides a (mol. wt 38000) and b (mol. wt 23000). The latter two polypeptides are supposedly fragments of α + ‐chains which have become partially cleaved by (auto)proteolysis.