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Purification of a phosphate carrier in pig heart mitochondria by affinity chromatography on mersalyl—ultrogel
Author(s) -
Touraille Sylvie,
Briand Yves,
Durand Rogert,
Bonnafous Jean-Claude,
Mani Jean-Claude
Publication year - 1981
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(81)81100-3
Subject(s) - chemistry , reprint , physics , astronomy
Mersalyl is a powerful inhibitor of phosphate transport in mitochondria. In addition, by its reversible reaction with thiol groups, it can protect the latter from irreversible reaction with N-ethylmaleimide [ 1,2]. Use of this property enabled a group of proteins of M, 3.0-3.2 X lo4 to be identified in the internal mitochondrial membrane [3-g]. This group of proteins could be directly labeled using radioactive mersalyl [6,9]. It was subsequently shown that these proteins consisted of a mersalyl and N-ethyhpaleimide-sensitive protein (M, 3.2 X 104) and the nucleotide translocase (M, 3.0 X IO”) [ IO,1 I]. These two proteins can be purified on hydroxyapatite [ 11 ,121. The mixture of the two proteins when integrated into liposomes catalyzes a Pi-Pi exchange which is sensitive to thiol-group reagents [ 121. This work describes the complete separation of the mersalyl and N-ethylmaleimide-sensitive protein from the nucleotide translocase. This purification was carried out by combining fractionation on a mersalylultrogel column with purification on hydroxylapatite.