Rapid quantitative characterization of protein interactions by composition gradient static light scattering.
Two new applications of the recently developed technique of composition gradient static light scattering (CG-SLS) are presented. 1), The method is demonstrated to be capable of detecting and quantitatively characterizing reversible association of chymotrypsin and bovine pancreatic trypsin inhibitor in a solution mixture and simultaneously occurring reversible self-association of chymotrypsin at low pH in the same mixture. The values of equilibrium constants for both self- and heteroassociation may be determined with reasonable precision from the analysis of data obtained from a single experiment requiring <15 min and <1 mg of each protein. 2), Analysis of the results of a single CG-SLS experiment carried out on Ftsz, a protein that self-associates to form linear oligomers of indefinite size in the presence of guanosine diphosphate, yields the dependence of the equilibrium constant for monomer addition upon oligomer size.
Kameyama
Keiichi
K
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, Maryland 20892-0830, USA.
Minton
Allen P
AP
D007611:Aprotinin / Q000032:analysis / Q000737:chemistry*; D002918:Chymotrypsin / Q000032:analysis / Q000737:chemistry*; D003199:Computer Systems; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods; D008027:Light; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D046912:Multiprotein Complexes / Q000032:analysis / Q000737:chemistry; D009391:Nephelometry and Turbidimetry / Q000295:instrumentation* / Q000379:methods; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000295:instrumentation* / Q000379:methods; D012542:Scattering, Radiation
D046912:Multiprotein Complexes; D007611:Aprotinin; D002918:Chymotrypsin