Abstract
1. A mol.wt. of 40030 +/- 830 has been estimated for phosphoglycerate kinase in concentrations less than 0.1 g/100 cm3 comparing favourably with expected values from X-ray diffraction measurements by 10% lower than the previously reported molecular weights made at higher concentrations. 2. The so20w, was estimated to be 3.12(+/-0.02)x10(-13)s and the coefficient had a low concentration dependency giving a g value (concentration-dependency) of 2.3 +/- 1.6cm3 .g-1. This agrees with previous qualitative observations. 3. By using fluctuation-intensity spectroscopy, the D20,w was estimated to be 7.4(+/-0.2)x10(-11)m2.s-1, and this was indistinguishable from the D20,w calculated from ultracentrifuge results. The water of hydration was estimated to be 0.46 g/g of protein. 4. It is inferred from the estimates that phosphoglycerate kinase associates with an interaction coefficient at 20 degrees C for monomer/dimer of between 10 and 12 cm3.g-1. 5. The ratio of molecular asymmetry (a/b) was estimated to be 2.5+/-0.2 from the values of D20,w and water of hydration. This compares favourably with the ratio from the overall dimensions estimated from X-ray diffraction measurements.
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Selected References
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- Adams M. J., Haas D. J., Jeffery B. A., McPherson A., Jr, Mermall H. L., Rossmann M. G., Schevitz R. W., Wonacott A. J. Low resolution study of crystalline L-lactate dehydrogenase. J Mol Biol. 1969 Apr;41(2):159–188. doi: 10.1016/0022-2836(69)90383-0. [DOI] [PubMed] [Google Scholar]
- BALDWIN R. L. Boundary spreading in sedimentation-velocity experiments. IV. Measurement of the standard deviation of a sedimentation-coefficient distribution: application to bovine albumin and beta-lactoglobulin. Biochem J. 1957 Mar;65(3):490–502. doi: 10.1042/bj0650490. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bryant T. N., Watson H. C., Wendell P. L. Structure of yeast phosphoglycerate kinase. Nature. 1974 Jan 4;247(5435):14–17. doi: 10.1038/247014a0. [DOI] [PubMed] [Google Scholar]
- Foord R., Jakeman E., Oliver C. J., Pike E. R., Blagrove R. J., Wood E., Peacocke A. R. Determination of diffusion coefficients of haemocyanin at low concentration by intensity fluctuation spectroscopy of scattered laser light. Nature. 1970 Jul 18;227(5255):242–245. doi: 10.1038/227242a0. [DOI] [PubMed] [Google Scholar]
- Gilbert L. M., Gilbert G. A. Sedimentation velocity measurement of protein association. Methods Enzymol. 1973;27:273–296. doi: 10.1016/s0076-6879(73)27014-3. [DOI] [PubMed] [Google Scholar]
- Kellett G. L. Dissociation of hemoglobin into subunits. Ligand-linked dissociation at neutral pH. J Mol Biol. 1971 Aug 14;59(3):401–424. doi: 10.1016/0022-2836(71)90307-x. [DOI] [PubMed] [Google Scholar]
- King T. A., Knox A., McAdam J. D. Translational and rotational diffusion of tobacco mosaic virus from polarized and depolarized light scattering. Biopolymers. 1973;12(8):1917–1926. doi: 10.1002/bip.1973.360120817. [DOI] [PubMed] [Google Scholar]
- Krietsch W. K., Bücher T. 3-phosphoglycerate kinase from rabbit sceletal muscle and yeast. Eur J Biochem. 1970 Dec;17(3):568–580. doi: 10.1111/j.1432-1033.1970.tb01202.x. [DOI] [PubMed] [Google Scholar]
- Kulbe K. D., Bojanovski M., Lamprecht W. Liver 3-phosphoglycerate kinase. Physico-chemical characterization of the bovine-liver enzyme. Eur J Biochem. 1975 Mar 17;52(2):239–254. doi: 10.1111/j.1432-1033.1975.tb03992.x. [DOI] [PubMed] [Google Scholar]
- Kuntz I. D. Hydration of macromolecules. IV. Polypeptide conformation in frozen solutions. J Am Chem Soc. 1971 Jan 27;93(2):516–518. doi: 10.1021/ja00731a037. [DOI] [PubMed] [Google Scholar]
- Kuntz I. D., Jr, Brassfield T. S., Law G. D., Purcell G. V. Hydration of macromolecules. Science. 1969 Mar 21;163(3873):1329–1331. doi: 10.1126/science.163.3873.1329. [DOI] [PubMed] [Google Scholar]
- Larsson-Raźnikiewicz M. Electrophoretic purification as well as some physical and chemical characterizations of phosphoglycerate kinase from yeast. Eur J Biochem. 1970 Sep;15(3):574–580. doi: 10.1111/j.1432-1033.1970.tb01043.x. [DOI] [PubMed] [Google Scholar]
- Raj T., Flygare W. H. Diffusion studies of bovine serum albumin by quasielastic light scattering. Biochemistry. 1974 Jul 30;13(16):3336–3340. doi: 10.1021/bi00713a024. [DOI] [PubMed] [Google Scholar]
- Scopes R. K. An improved procedure for the isolation of 3-phosphoglycerate kinase from yeast. Biochem J. 1971 Mar;122(1):89–92. doi: 10.1042/bj1220089. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spragg S. P. Processing ultracentrifuge data with an "on-line" digital computer. Anal Chim Acta. 1967 May-Jun;38(1):137–142. doi: 10.1016/s0003-2670(01)80569-9. [DOI] [PubMed] [Google Scholar]
