High-level bacterial expression of mitochondrial transport proteins.
The inability to obtain abundant quantities of purified, functional membrane proteins has represented a significant impediment to the goal of obtaining high-resolution structural information. In this review, procedures are described which have been developed in this laboratory and enable the high-level bacterial expression and subsequent purification of functional mitochondrial citrate transport proteins from yeast and rat liver. The data that we have obtained using these procedures and related results from other laboratories are discussed. Additionally, the general applicability of this approach to most mitochondrial transport proteins as well as to other types of membrane proteins is considered. Finally, relevant considerations when contemplating the use of this methodology and the likely value of this approach in future areas of research are explored.
Kaplan
R S
RS
Department of Pharmacology, College of Medicine, University of South Alabama, Mobile 36688, USA.
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification; D002951:Citrates / Q000378:metabolism; D019343:Citric Acid; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D008099:Liver / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
D002352:Carrier Proteins; D002951:Citrates; D011994:Recombinant Proteins; C020723:citrate-binding transport protein; D019343:Citric Acid