Purification and characterization of a heavy-metal-modulated nuclear protein from SV40-transformed cells

Abstract

Sodium arsenite was found to stimulate an SV40-transformed BALB/c cell line (SVT2) to synthesize a 31-kDa protein within 2.5 h. This SVT2 protein was purified to homogeneity. It is a nuclear protein which appears to be associated with membranes because it is not extractable from nuclear membrane preparations by 2 M salt. It is highly hydrophobic, eluting from a reverse-phase HPLC column at a similar acetonitrile concentration as a previously described 31-kDa BALB/c-3T3 cell nuclear protein. However, digestion of highly purified BALB/c-3T3 and SVT2 cell proteins with V8 protease revealed nonidentical fragmentation patterns. Moreover, amino acid analysis of the two proteins was also dissimilar, indicating different primary structures. Thus, these two nuclear membrane associated proteins appear to be distinct species.