Pi binding by the F1-ATPase of beef heart mitochondria and of the Escherichia coli plasma membrane.

Abstract:

:Pi binding by the F(1)-ATPase of beef heart mitochondria and of the Escherichia coli plasma membrane (E. coli F(1)) was examined by two methods: the centrifuge column procedure [Penefsky, H.S. (1977) J. Biol. Chem. 252, 2891-2899] and the Paulus pressure dialysis cell [Paulus, H. (1969) Anal. Biochem. 32, 91-100]. The latter is an equilibrium dialysis-type procedure. Pi binding by beef heart F(1) could be determined by either procedure. However, direct binding of Pi to E. coli F(1) could be determined adequately only in the Paulus cell which indicated more than two binding sites per mol of enzyme with a K(d) in the range of 0.1 mM. It is concluded that previous failure to observe Pi binding to E. coli F(1) with the centrifuge column procedure is due to a rapid rate of dissociation of Pi from the E. coli enzyme which results in loss of Pi during transit of the enzyme-Pi complex through the column.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Penefsky HS

doi

10.1016/j.febslet.2005.02.072

keywords:

subject

Has Abstract

pub_date

2005-04-11 00:00:00

pages

2250-2

issue

10

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(05)00330-3

journal_volume

579

pub_type

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