The 46-kDa nucleoside triphosphatase of rat liver nuclear scaffold represents the N-terminal portion of lamins A/C.
The major nucleoside triphosphatase (NTPase) of rat liver nuclear scaffold (NS) or envelope, which is thought to participate in nucleocytoplasmic transport, has been identified via photoaffinity labeling as a 46-kDa polypeptide. This 46-kDa protein was purified by SDS-polyacrylamide gel electrophoresis and cleaved with trypsin. The resulting peptides were purified by HPLC and five were microsequenced. All five peptides appear to be derived from the N-terminal region of lamins A/C. Subsequent experiments with photolabeled NS showed that the 46-kDa polypeptide was selectively immunoprecipitated by antiserum specific to lamins A/C and by affinity-purified anti-lamin antibodies. Photolabeling of nuclei prepared in the presence of protease inhibitors showed predominant labeling of the 46-kDa polypeptide, suggesting that it is an integral nuclear constituent and not an artifact produced during NS preparation. Use of protease inhibitors throughout purification of NS increased the specificity of photolabeling of the 46-kDa band by significantly reducing photolabeling of smaller molecular weight components, which arise by proteolysis. Anti-lamin antibodies also produced a significant inhibition of NTPase activity in NS. These results suggest that the N-terminal portion of lamins A/C represents the 46-kDa NTPase, which, according to previous reports, may participate in RNA transport.
Clawson
G A
GA
Department of Pathology, School of Medicine, University of California, San Francisco 94143.
Lackey
A
A
Tokes
Z A
ZA
D000345:Affinity Labels; D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D034882:Lamins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins; D043583:Nucleoside-Triphosphatase; D010744:Phosphoric Monoester Hydrolases; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats
D000345:Affinity Labels; D034882:Lamins; D009687:Nuclear Proteins; D011480:Protease Inhibitors; D010744:Phosphoric Monoester Hydrolases; D043583:Nucleoside-Triphosphatase