A comparison of native and covalently crosslinked creatine kinases: denaturation and reassembly

Archives of Biochemistry and Biophysics
S H GrossmanD Mixon

Abstract

Creatine kinase from rabbit muscle, a dimeric protein of molecular weight 81,000, was crosslinked with glutaraldehyde. The time course of the reaction was monitored by inhibition of activity, fluorescence polarization, and electrophoretic mobility in polyacrylamide slab gels containing sodium dodecyl sulfate. Inactivation is complete in 40 min. ADP (1.2 mM) decreases the rate but does not prevent complete inactivation. Fluorescence polarization analysis of N-dansylaziridyl creatine kinase suggests the formation of high molecular weight oligomers. This is confirmed through electrophoretic molecular weight analysis, which demonstrates the formation of products with molecular weight greater than 150,000 throughout the course of reaction with glutaraldehyde. During crosslinking at least three other products appear with apparent molecular weights of 150,000, 95,000, and 85,000. The 150,000 and 95,000 molecular weight products are transient, disappearing after 40 min of exposure to glutaraldehyde. Monomers of 40,000 molecular weight have also been eliminated by 40 min. The 85,000 molecular weight product, which is not present until 10 min after crosslinking, is the only product other than oligomers present at 60 min of crosslinking. ...Continue Reading

References

May 1, 1976·FEBS Letters·G F Bickerstaff, N C Price
Dec 29, 1974·Biochimica Et Biophysica Acta·I Ramasamy, P J Butterworth
Jul 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·G E Davies, G R Stark
Apr 28, 1968·Journal of Molecular Biology·W H Bishop, F M Richards
Nov 22, 1965·Biochemical and Biophysical Research Communications·D M DawsonN O Kaplan
Sep 1, 1964·Proceedings of the National Academy of Sciences of the United States of America·F A QUIOCHO, F M RICHARDS

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Citations

Feb 1, 1985·Archives of Biochemistry and Biophysics·S H Grossman, D Mixon
Aug 10, 2002·Journal of Bacteriology·Daniel C ScottPhillip E Klebba
Jul 1, 1986·Archives of Biochemistry and Biophysics·S H GrossmanJ J Lense

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