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
[1]   SEQUENCES REQUIRED FOR DELIVERY AND LOCALIZATION OF THE ADP/ATP TRANSLOCATOR TO THE MITOCHONDRIAL INNER MEMBRANE [J].
ADRIAN, GS ;
MCCAMMON, MT ;
MONTGOMERY, DL ;
DOUGLAS, MG .
MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (02) :626-634
[2]   ASYMMETRIC ORIENTATION OF THE RECONSTITUTED ASPARTATE GLUTAMATE CARRIER FROM MITOCHONDRIA [J].
DIERKS, T ;
KRAMER, R .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 937 (01) :112-126
[3]   ABUNDANT BACTERIAL EXPRESSION AND RECONSTITUTION OF AN INTRINSIC MEMBRANE-TRANSPORT PROTEIN FROM BOVINE MITOCHONDRIA [J].
FIERMONTE, G ;
WALKER, JE ;
PALMIERI, F .
BIOCHEMICAL JOURNAL, 1993, 294 :293-299
[4]   IDENTIFICATION AND PURIFICATION OF THE CARNITINE CARRIER FROM RAT-LIVER MITOCHONDRIA [J].
INDIVERI, C ;
TONAZZI, A ;
PALMIERI, F .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1020 (01) :81-86
[5]   Identification of a novel gene encoding the yeast mitochondrial dicarboxylate transport protein via overexpression, purification, and characterization of its protein product [J].
Kakhniashvili, D ;
Mayor, JA ;
Gremse, DA ;
Xu, Y ;
Kaplan, RS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (07) :4516-4521
[6]  
Kaplan RS, 1996, J BIOENERG BIOMEMBR, V28, P41
[7]   STREPTOZOTOCIN-INDUCED ALTERATIONS IN THE LEVELS OF FUNCTIONAL MITOCHONDRIAL ANION TRANSPORT PROTEINS [J].
KAPLAN, RS ;
OLIVEIRA, DL ;
WILSON, GL .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1990, 280 (01) :181-191
[8]  
KAPLAN RS, 1993, J BIOL CHEM, V268, P13682
[9]  
KAPLAN RS, 1990, J BIOL CHEM, V265, P13379
[10]   HIGH-LEVEL EXPRESSION AND CHARACTERIZATION OF THE MITOCHONDRIAL CITRATE TRANSPORT PROTEIN FROM THE YEAST SACCHAROMYCES-CEREVISIAE [J].
KAPLAN, RS ;
MAYOR, JA ;
GREMSE, DA ;
WOOD, DO .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (08) :4108-4114