Static light scattering studies of OmpF porin: implications for integral membrane protein crystallization.
Integral membrane proteins carry out some of the most important functions of living cells, yet relatively few details are known about their structures. This is due, in large part, to the difficulties associated with preparing membrane protein crystals suitable for X-ray diffraction analysis. Mechanistic studies of membrane protein crystallization may provide insights that will aid in determining future membrane protein structures. Accordingly, the solution behavior of the bacterial outer membrane protein OmpF porin was studied by static light scattering under conditions favorable for crystal growth. The second osmotic virial coefficient (B22) was found to be a predictor of the crystallization behavior of porin, as has previously been found for soluble proteins. Both tetragonal and trigonal porin crystals were found to form only within a narrow window of B22 values located at approximately -0.5 to -2 X 10(-4) mol mL g(-2), which is similar to the "crystallization slot" observed for soluble proteins. The B22 behavior of protein-free detergent micelles proved very similar to that of porin-detergent complexes, suggesting that the detergent's contribution dominates the behavior of protein-detergent complexes under crystallizing conditions. This observation implies that, for any given detergent, it may be possible to construct membrane protein crystallization screens of general utility by manipulating the solution properties so as to drive detergent B22 values into the crystallization slot. Such screens would limit the screening effort to the detergent systems most likely to yield crystals, thereby minimizing protein requirements and improving productivity.
Hitscherich
C
C
Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City 52242, USA.
Kaplan
J
J
Allaman
M
M
Wiencek
J
J
Loll
P J
PJ
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D003460:Crystallization; D003461:Crystallography; D003902:Detergents / Q000737:chemistry; D003956:Dialysis; D004926:Escherichia coli / Q000201:enzymology; D008027:Light; D008823:Micelles; D008958:Models, Molecular; D011092:Polyethylene Glycols / Q000737:chemistry; D018272:Porins / Q000737:chemistry*; D011487:Protein Conformation; D012542:Scattering, Radiation; D012984:Software; D014961:X-Ray Diffraction
D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D008823:Micelles; C109237:OmpF protein; D018272:Porins; D011092:Polyethylene Glycols