Size determination of multienzyme complexes using two-dimensional agarose gel electrophoresis

Proteins
R A EasomP Serwer

Abstract

In studies of the size and structure of multienzyme complexes, a procedure complementary to electron microscopy for determining the molecular dimensions of hydrated multisubunit complexes is needed. For some applications this procedure must be capable of detecting aggregation of complexes and must be applicable to impure preparations. In the present study, a procedure of two-dimensional agarose gel electrophoresis (2d-AGE) (Serwer, P. et al. Anal. Biochem. 152:339-345, 1986) was modified and employed to provide accurate size measurements of several classical multienzyme complexes. To improve band clarity and to achieve required gel pore sizes, a hydroxyethylated agarose was used. The effective pore's radius (PE) as a function of gel concentration was determined for this agarose in the range of PE values needed for multienzyme complexes (effective radius, R = 10-30 nm). Appropriate conditions were established to measure R values +/- 1% of the pyruvate (PDC), alpha-ketoglutarate (alpha-KGDC), and the branched chain alpha-keto acid (BCDC) dehydrogenase multienzyme complexes; the accuracy of R was limited by the accuracy of the determinations of the R value for the size standards. The PDC from bovine heart was found to have an R = ...Continue Reading

References

Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D M BleileL J Reed
Mar 1, 1976·International Journal of Radiation Oncology, Biology, Physics·S B FieldR Morrison
Jan 1, 1975·Annual Review of Biophysics and Bioengineering·D M Engelman, P B Moore
Feb 1, 1986·Analytical Biochemistry·P SerwerG A Griess
Apr 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·D Rodbard, A Chrambach
Sep 15, 1973·Journal of Molecular Biology·F W Studier
Oct 14, 1968·Journal of Molecular Biology·F M Richards, J R Knowles
Mar 1, 1982·Journal of Autonomic Pharmacology·S A Doggrell, L Vincent
Jul 3, 1980·Nature·C Abad-ZapateroT Tsukihara

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Citations

May 1, 1995·The International Journal of Biochemistry & Cell Biology·T Pawelczyk, M S Olson
Jul 19, 1994·Proceedings of the National Academy of Sciences of the United States of America·S D Jett, D G Bear
Apr 1, 1992·Archives of Biochemistry and Biophysics·T PawelczykM S Olson
Apr 28, 1995·Journal of Chromatography. a·P SerwerG A Griess

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