Document Detail


Functionally important conserved length of C-terminal regions of yeast and bovine ADP/ATP carriers, identified by deletion mutants studies, and water accessibility of the amino acids at the C-terminal region of the yeast carrier.
MedLine Citation:
PMID:  18313366     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Comparison of the amino acid sequence of yeast type 2 ADP/ATP carrier (yAAC2) with that of bovine type 1 AAC (bAAC1) revealed that the N- and C-terminus of yAAC2 are 15- and 6-amino acids longer, respectively, than those of bAAC1. In the present study, we focused on the difference in the C-terminal region between yAAC2 and bAAC1. Deletion of first six residues of C-terminus of yAAC did not markedly affect the function of yAAC2; however, further deletion of 1 amino acid (7th amino acid from the C-terminus) destroyed its function. On the contrary, deletion of the first amino acid residue of the C-terminus of bAAC1 caused failure of its functional expression in yeast mitochondria. Based on these results, we concluded that the 6-amino acid residue extension of the C-terminus of yAAC2 was not necessary for the function of this carrier and that the remainder of the C-terminal region of yAAC2, having a length conserved with that of bAAC1, is important for the transport function of AACs. We next prepared various single-Cys mutants in which each of 32 residues in the C-terminus of yAAC2 was replaced by a Cys residue. Since all mutants were successfully expressed in yeast mitochondria, we examined the reactivity of these cysteine residues with the membrane-impermeable sulfhydryl reagent eosin 5-maleimide (EMA). As a result, all cysteine residues that replaced the 9 continuous amino acids in Met310-Lys318 showed high reactivity with EMA regardless of the presence of carboxyatractyloside or bongkrekic acid; and so this region was concluded to be exposed to the water-accessible environment. Furthermore, based on the reactivities of cysteine residues that replaced amino acids in the sixth transmembrane segment, the probable structural features of the C-terminal region of this carrier in the presence of bongkrekic acid were discussed.
Authors:
Akihiro Iwahashi; Aoi Ishii; Naoshi Yamazaki; Mitsuru Hashimoto; Kazuto Ohkura; Masatoshi Kataoka; Eiji Majima; Hiroshi Terada; Yasuo Shinohara
Related Documents :
10373116 - Preventing neurodegeneration in the drosophila mutant bubblegum.
17029416 - Structure/function mapping of amino acids in the n-terminal zinc finger of the human im...
10091326 - Cloning of the transketolase gene and the effect of its dosage on aromatic amino acid p...
2227406 - Branched chain amino acid regulation of the ilv2 locus in saccharomyces cerevisiae.
1439206 - Effects of oral sunflower oil and olive oil on serum and cutaneous fatty acid concentra...
17410406 - Relationship between (na + k)-atpase activity, lipid peroxidation and fatty acid profil...
Publication Detail:
Type:  Comparative Study; Journal Article     Date:  2008-02-05
Journal Detail:
Title:  Mitochondrion     Volume:  8     ISSN:  1567-7249     ISO Abbreviation:  Mitochondrion     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-03-10     Completed Date:  2008-07-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100968751     Medline TA:  Mitochondrion     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  196-204     Citation Subset:  IM    
Affiliation:
Institute for Genome Research, University of Tokushima, Kuramotocho-3, Tokushima 770-8503, Japan.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Bongkrekic Acid / pharmacology
Cattle
Eosine Yellowish-(YS) / analogs & derivatives,  metabolism
Gene Deletion
Mitochondrial ADP, ATP Translocases / drug effects,  genetics*,  metabolism
Molecular Sequence Data
Protein Conformation
Saccharomyces cerevisiae Proteins / drug effects,  genetics*,  metabolism
Sequence Alignment
Transformation, Genetic
Chemical
Reg. No./Substance:
0/PET9 protein, S cerevisiae; 0/Saccharomyces cerevisiae Proteins; 11076-19-0/Bongkrekic Acid; 17372-87-1/Eosine Yellowish-(YS); 76296-42-9/eosin maleimide; 9068-80-8/Mitochondrial ADP, ATP Translocases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Estimating novel potential drug targets of Plasmodium falciparum by analysing the metabolic network ...
Next Document:  Systematic evaluation of muscle coenzyme Q10 content in children with mitochondrial respiratory chai...