Bacterial overexpression of putative yeast mitochondrial transport proteins

被引:9
作者
Mayor, JA
Kakhniashvili, D
Gremse, DA
Campbell, C
Krämer, R
Schroers, A
Kaplan, RS
机构
[1] Finch Univ Hlth Sci Chicago Med Sch, Dept Biol Chem, N Chicago, IL 60064 USA
[2] Univ S Alabama, Coll Med, Dept Pharmacol, Mobile, AL 36688 USA
[3] Univ Cologne, Inst Biochem, D-50674 Cologne, Germany
关键词
mitochondria; transporter; overexpression; yeast; membrane proteins; crystallization;
D O I
10.1023/A:1022426900735
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Thirty-two genes have been identified within the genome of the yeast Saccharomyces cerevisiae which putatively encode mitochondrial transport proteins. We have attempted to overexpress a subset of these genes, namely those which encode mitochondrial transporters of unknown function, and have succeeded in overexpressing 19 of these genes. The overexpressed proteins were then isolated and tested for five well-characterized reconstituted transport activities (i.e., the transport of citrate, dicarboxylates, pyruvate, carnitine, and aspartate). Utilizing this approach, we have clearly identified the yeast mitochondrial dicarboxylate transport protein, as well as two additional lower-magnitude transport functions (i.e., tricarboxylate and dicarboxylate transport activities). The implications of these results and the considerations relevant to this approach are discussed.
引用
收藏
页码:541 / 547
页数:7
相关论文
共 19 条
[11]  
KAPLAN RS, 1996, MOL BIOL MEMBRANE TR, P277
[12]  
KOLAROV J, 1990, J BIOL CHEM, V265, P12711
[13]   FUNCTIONAL RECONSTITUTION OF CARRIER PROTEINS BY REMOVAL OF DETERGENT WITH A HYDROPHOBIC ION-EXCHANGE COLUMN [J].
KRAMER, R ;
HEBERGER, C .
BIOCHIMICA ET BIOPHYSICA ACTA, 1986, 863 (02) :289-296
[14]  
LAWSON JE, 1988, J BIOL CHEM, V263, P14812
[15]   KINETIC STUDY OF TRICARBOXYLATE CARRIER IN RAT-LIVER MITOCHONDRIA [J].
PALMIERI, F ;
QUAGLIARIELLO, E ;
STIPANI, I ;
KLINGENBERG, M .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1972, 26 (04) :587-+
[16]   Identification by bacterial expression and functional reconstitution of the yeast genomic sequence encoding the mitochondrial dicarboxylate carrier protein [J].
Palmieri, L ;
Palmieri, F ;
Runswick, MJ ;
Walker, JE .
FEBS LETTERS, 1996, 399 (03) :299-302
[17]   Identification of the yeast ARG-11 gene as a mitochondrial ornithine carrier involved in arginine biosynthesis [J].
Palmieri, L ;
DeMarco, V ;
Iacobazzi, V ;
Palmieri, F ;
Runswick, MJ ;
Walker, JE .
FEBS LETTERS, 1997, 410 (2-3) :447-451
[18]   FLX1 codes for a carrier protein involved in maintaining a proper balance of flavin nucleotides in yeast mitochondria [J].
Tzagoloff, A ;
Jang, J ;
Glerum, DM ;
Wu, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (13) :7392-7397
[19]   YEAST MITOCHONDRIAL PHOSPHATE-TRANSPORT PROTEIN EXPRESSED IN ESCHERICHIA-COLI - SITE-DIRECTED MUTATIONS AT THREONINE-43 AND AT A SIMILAR LOCATION IN THE 2ND TANDEM REPEAT (ISOLEUCINE-141) [J].
WOHLRAB, H ;
BRIGGS, C .
BIOCHEMISTRY, 1994, 33 (32) :9371-9375