High resolution KSCN/CsSCN equilibrium gradients effectively separate a population of density labeled proteins from unlabeled proteins

Analytical Biochemistry
W H PhillipsR M Grainger

Abstract

The persistence of proteins in a number of biological systems has been analyzed by density labeling techniques; however, the utility of this approach has been severely hampered by poor resolution between density-labeled and unlabeled proteins on equilibrium gradients. A high resolution equilibrium salt gradient composed of KSCN/CsSCN has been developed to effectively separate density-labeled proteins (13C-15N-2H-substituted) from unlabeled proteins. The resolution of this system is approximately twofold greater than that previously achieved with cesium formate/guanidine hydrochloride equilibrium gradients which have been used in many recent protein density labeling studies. In order to examine the extent of cross-contamination between density-labeled and unlabeled proteins in a KSCN/CsSCN gradient system, density-labeled chick epidermal proteins were mixed with unlabeled Drosophila larval proteins and then separated on these equilibrium gradients. From individual gradient fractions proteins were recovered and fractionated on a sodium dodecyl sulfate-polyacrylamide gel, demonstrating the virtually complete separation between the two populations. The general utility of this system for protein stability studies is also demonstrated.

References

Aug 15, 1975·European Journal of Biochemistry·R A Laskey, A D Mills
Nov 1, 1975·The Biochemical Journal·A BoudetD D Davies
Nov 1, 1985·The Journal of Cell Biology·J A Winkles, R M Grainger
Dec 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·R M Grainger, N K Wessells
Jul 1, 1974·European Journal of Biochemistry·W M Bonner, R A Laskey
Aug 14, 1969·Journal of Molecular Biology·L S Williams, F C Neidhardt
May 11, 1983·Nucleic Acids Research·I M Leffak
May 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·V P KnutsonM D Lane
Jul 15, 1958·Proceedings of the National Academy of Sciences of the United States of America·M Meselson, F W Stahl

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