Site-directed mutagenesis of the yeast mitochondrial ADP/ATP translocator: Six arginines and one lysine are essential

David Nelson, Janet E. Lawson, Martin Klingenberg, Michael G. Douglas

Research output: Contribution to journalArticle

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Abstract

The ADP/ATP translocator mediates adenine nucleotide exchange across the inner mitochondrial membrane. ADP/ATP exchange is essential when yeast are grown on a non-fermentable carbon source such as glycerol, but it is not required for growth on glucose. Failure to grow on glycerol is therefore a phenotypic indicator of protein function, and it has been used here to screen site-directed mutants to identify functionally important amino acids in the yeast adenine nucleotide translocator (AAC2). Single mutations of all four charged amino acids in the transmembrane segments of AAC2 (K38A, R96D, R96H, R96L, R96P, R204L, R294A) resulted in loss of function, as did mutations in the matrix arginine cluster (R252I, R253I, R254I). Seven other residues were mutated without affecting growth on glycerol (C73S, C244S, C271S, K179M, K182I, P247G, W235F). The non-functional mutants have been used to select intragenic suppressors to gain further insight into the structure of this membrane transport protein.

Original languageEnglish (US)
Pages (from-to)1159-1170
Number of pages12
JournalJournal of Molecular Biology
Volume230
Issue number4
DOIs
StatePublished - Jan 1 1993

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Site-Directed Mutagenesis
Glycerol
Adenosine Diphosphate
Lysine
Arginine
Adenine Nucleotides
Adenosine Triphosphate
Yeasts
Amino Acids
Mutation
Membrane Transport Proteins
Mitochondrial Membranes
Growth
Carbon
Glucose
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology

Cite this

Site-directed mutagenesis of the yeast mitochondrial ADP/ATP translocator : Six arginines and one lysine are essential. / Nelson, David; Lawson, Janet E.; Klingenberg, Martin; Douglas, Michael G.

In: Journal of Molecular Biology, Vol. 230, No. 4, 01.01.1993, p. 1159-1170.

Research output: Contribution to journalArticle

Nelson, David ; Lawson, Janet E. ; Klingenberg, Martin ; Douglas, Michael G. / Site-directed mutagenesis of the yeast mitochondrial ADP/ATP translocator : Six arginines and one lysine are essential. In: Journal of Molecular Biology. 1993 ; Vol. 230, No. 4. pp. 1159-1170.
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