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Journal of Bacteriology, June 1999, p. 3688-3694, Vol. 181, No. 12
Max-Planck-Institut für Biologie,
Abteilung Mikrobiologie, D-72076 Tübingen, Germany
Received 25 January 1999/Accepted 21 April 1999
The N-terminal domain of the OmpA protein from Escherichia
coli, consisting of 170 amino acid residues, is embedded in the outer membrane, in the form of an antiparallel
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Copyright © 1999, American Society for Microbiology. All rights reserved.
Structural and Functional Roles of the
Surface-Exposed Loops of the
-Barrel Membrane Protein OmpA from
Escherichia coli
-barrel whose eight
transmembrane
-strands are connected by three short periplasmic turns and four relatively large surface-exposed hydrophilic loops. This
protein domain serves as a paradigm for the study of membrane assembly
of integral
-structured membrane proteins. In order to dissect the
structural and functional roles of the surface-exposed loops, they were
shortened separately and in all possible combinations. All 16 loop
deletion mutants assembled into the outer membrane with high efficiency
and adopted the wild-type membrane topology. This systematic approach
proves the absence of topogenic signals (e.g., in the form of loop
sizes or charge distributions) in these loops. The shortening of
surface-exposed loops did not reduce the thermal stability of the
protein. However, none of the mutant proteins, with the exception of
the variant with the fourth loop shortened, served as a receptor for
the OmpA-specific bacteriophage K3. Furthermore, all loops were
necessary for the OmpA protein to function in the stabilization of
mating aggregates during F conjugation. An OmpA deletion variant with
all four loops shortened, consisting of only 135 amino acid residues,
constitutes the smallest
-structured integral membrane protein known
to date. These results represent a further step toward the development
of artificial outer membrane proteins.
*
Present address: Martin-Luther-University, Institute of
Genetics, Weinbergweg 22, D-06120 Halle (Saale), Germany. Phone:
49-345-5526293. Fax: 49-345-5527259. E-mail:
koebnik{at}iname.com.
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