NDLI: Properties and purification of an active biotinylated lactose permease from Escherichia coli.

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Properties and purification of an active biotinylated lactose permease from Escherichia coli.

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Properties and purification of an active biotinylated lactose permease from Escherichia coli.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Privé, G. G. Consler, T. G. Kaback, H. R. Verner, G. E. Zen, K. H. Jung, H. Persson, B. L. Jung, K.
Description	Author Affiliation: Consler TG ( Howard Hughes Medical Institute, Department of Physiology, University of California, Los Angeles 90024-1662.);
Abstract	A simplified approach for purification of functional lactose permease from Escherichia coli is described that is based on the construction of chimeras between the permease and a 100-amino acid residue polypeptide containing the biotin acceptor domain from the oxaloacetate decarboxylase of Klebsiella pneumoniae [Cronan, J. E., Jr. (1990) J. Biol. Chem. 265, 10327-10333]. Chimeras were constructed with a factor Xa protease site and the biotin acceptor domain in the middle cytoplasmic loop (loop 6) or at the C terminus of the permease. Each construct catalyzes active lactose transport in cells and right-side-out membrane vesicles. Moreover, the constructs are biotinylated in vivo, and in both chimeras, the factor Xa protease site is accessible from the cytoplasmic surface of the membrane. Both biotinylated permeases bind selectively to immobilized monomeric avidin and are eluted with free biotin in a high state of purity, and the loop 6 chimera catalyzes active transport after reconstitution into proteoliposomes. The methodology described should be applicable to other membrane proteins.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	15
Volume Number	90
Language	English
Publisher	National Academy of Sciences
Publisher Date	1993-09-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Escherichia Coli Proteins Membrane Transport Proteins Isolation & Purification Monosaccharide Transport Proteins Symporters Bacterial Proteins Biological Transport Biotin Chromatography, Affinity Escherichia Coli Chemistry Lactose Metabolism Membrane Proteins Recombinant Fusion Proteins Research Support, Non-U.S. Gov't Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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