Outer membrane protein A of E. coli folds into detergent micelles, but not in the presence of monomeric detergent

Protein Science - Tập 8 Số 10 - Trang 2065-2071 - 1999
Jörg H. Kleinschmidt1, Michael C. Wiener2, Lukas K. Tamm2
1Department of Molecular Physiology and Biological Physics and Center for Structural Biology, University of Virginia Health Sciences Center, Charlottesville 22906-0011, USA.
2Department of Molecular Physiology and Biological Physics, and Center for Structural Biology, University of Virginia Health Sciences Center, P.O. Box 10011, Charlottesville, Virginia 22906-0011

Tóm tắt

AbstractOuter membrane protein A (OmpA) of Escherichia coli is a β‐barrel membrane protein that unfolds in 8 M urea to a random coil. OmpA refolds upon urea dilution in the presence of certain detergents or lipids. To examine the minimal requirements for secondary and tertiary structure formation in β‐barrel membrane proteins, folding of OmpA was studied as a function of the hydrophobic chain length, the chemical structure of the polar headgroup, and the concentration of a large array of amphiphiles. OmpA folded in the presence of detergents only above a critical minimal chain length of the apolar chain as determined by circular dichroism spectroscopy and a SDS‐PAGE assay that measures tertiary structure formation. Details of the chemical structure of the polar headgroup were unimportant for folding. The minimal chain length required for folding correlated with the critical micelle concentration in each detergent series. Therefore, OmpA requires preformed detergent micelles for folding and does not adsorb monomeric detergent to its perimeter after folding. Formation of secondary and tertiary structure is thermodynamically coupled and strictly dependent on the interaction with aggregated amphiphiles.

Từ khóa


Tài liệu tham khảo

Bailyes EM, 1983, Solubilization and purification of rat liver 5′‐nucleotidase by use of a zwitterionic detergent and a monoclonal‐antibody immunoadsorbent, Biochem J, 203, 243

10.1016/0165-022X(80)90044-5

Cevc G, 1987, Phospholipid bilayers. Physical principles and models.

10.1016/S0021-9258(17)30602-6

Eisele JL, 1990, <123>In vitro</123> folding and oligomerization of a membrane protein. Transition of bacterial porin from random coil to native conformation, J Biol Chem, 265, 10217, 10.1016/S0021-9258(18)86933-2

10.1016/0003-2697(78)90565-1

10.1016/0076-6879(79)56066-2

10.1016/S0021-9258(19)45190-9

Helenius A, 1975, Solubilization of membranes by detergents, Biochim Biophys Acta, 415, 133, 10.1016/0304-4157(75)90016-7

10.1021/bi982465w

10.1021/bi961478b

10.1111/j.1462-5822.2007.00901.x

10.1111/j.1432-1033.1983.tb07080.x

Lund S, 1989, Detergent structure and associated lipid as determinants in the stabilization of solubilized Ca2+‐ATPase from sarcoplasmic reticulum, J Biol Chem, 264, 4907, 10.1016/S0021-9258(18)83677-8

Makino S, 1973, The binding of deoxycholate and Triton X‐100 to proteins, J Biol Chem, 248, 4926, 10.1016/S0021-9258(19)43653-3

Makino S, 1975, Interaction of deoxycholate and of detergents with the coat protein of bacteriophage, J Biol Chem, 250, 4327, 10.1016/S0021-9258(19)41422-1

10.1016/S0021-9258(17)46681-6

10.1016/S0021-9258(19)42440-X

10.1038/2983

10.1021/bi00673a025

Rosen MJ, 1989, Surfactants and interfacial phenomena

10.1016/1047-8477(90)90068-N

10.1111/j.1432-1033.1978.tb12013.x

Shinoda K., 1978, Principles of solution and solubility.

10.1016/S0076-6879(97)76050-6

Sugarawa E, 1996, Secondary structure of the outer membrane proteins OmpA of <123>Escherichia coli</123> and OprF of Pseudomonas aeruginosa, J Bacteriol, 178, 6067, 10.1128/jb.178.20.6067-6069.1996

10.1073/pnas.89.16.7457

Tanford C., 1980, The hydrophobic effect: Formation of micelles and biological membranes

10.1016/0304-4157(76)90009-5

10.1016/0301-4622(74)80004-9

10.1016/0022-2836(86)90292-5

Weber K, 1964, The reliability of molecular weight determinations by dodecyl sulfate‐polyacrylamide gel electrophoresis, J Biol Chem, 244, 4406, 10.1016/S0021-9258(18)94333-4

Zulauf M., 1990, Crystallization of membrane proteins., 53

Thông tin
Thông tin xuất bản

Protein Science

Tập 8 Số 10

2065-2071

Thông tin tác giả