The ADP/ATP carrier from yeast (AAC-2) is uniquely suited for the assignment of the binding center by photoaffinity labeling

Peter Mayinger, Edith Winkler, Martin Klingenberg

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The ADP/ATP carrier from yeast was photoaffinity-labeled in mitochondria with 2-azido-[α-32P]ATP in a binding-center-specific, i.e. carboxyatractylate-sensitive, manner. After isolation, fragmentation possibilities unique for the yeast AAC-2 could be exploited to assign the insertion to a narrow range of the sequence. The CNBr fragment 115-210 contained all the incorporated label which corresponds to the second domain within the triple-domain primary structure of the AAC. With hydroxylamine cleavage directed to the Asn 171-Gly 172 site, all the label was found in the C-terminal 16 kDa fragment. Thus the 2-azido-ATP incorporation is clearly delimited to the 172-210 segment. 8-Azido-[α-32P]ATP could be site-specifically incorporated only in isolated AAC since it has a much lower affinity for AAC than 2-azido-ATP. The label was also exclusively found in the 172-210 region. With both forms no incorporation into the C-terminal region was found, as claimed for bovine AAC. The labeled segment contains Lys 179 and 182 which are homologous to bovine Lys 162 and 165 and which have been proposed to be in the translocation path.

Original languageEnglish (US)
Pages (from-to)421-426
Number of pages6
JournalFEBS Letters
Volume244
Issue number2
DOIs
StatePublished - Feb 27 1989
Externally publishedYes

Fingerprint

aminoglycoside 2'-N-acetyltransferase
ATP Translocases Mitochondrial ADP
Yeast
Labeling
Labels
Yeasts
Adenosine Triphosphate
Hydroxylamine
Mitochondria
2-azidoadenosine 5'-triphosphate

Keywords

  • (Saccharomyces cerevisiae)
  • ADP/ATP carrier
  • Azido-ATP, 2-
  • Azido-ATP, 8-
  • Photoaffinity labeling

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

The ADP/ATP carrier from yeast (AAC-2) is uniquely suited for the assignment of the binding center by photoaffinity labeling. / Mayinger, Peter; Winkler, Edith; Klingenberg, Martin.

In: FEBS Letters, Vol. 244, No. 2, 27.02.1989, p. 421-426.

Research output: Contribution to journalArticle

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