PMID: 9540798Apr 16, 1998Paper

Interactions of serine proteinases with pNiXa, a serpin of Xenopus oocytes and embryos

Biochimica Et Biophysica Acta
J KotyzaF W Sunderman

Abstract

In a previous study, kinetic assays showed that pNiXa, an Ni(II)-binding serpin of Xenopus oocytes and embryos, strongly inhibits bovine chymotrypsin, weakly inhibits porcine elastase, and does not inhibit bovine trypsin. In this study, analyses by SDS-PAGE and gelatin zymography showed that an SDS-resistant complex is formed upon the interaction of pNiXa with bovine chymotrypsin. No such pNiXa-enzyme complex was detected after pNiXa interactions with porcine elastase, bovine trypsin, or human cathepsin G. The major products of pNiXa cleavage by the four proteinases were partially sequenced by Edman degradation. The cleavage products were also tested by immunoblotting with an antibody to the His-cluster of pNiXa, and by radio-blotting with 63Ni(II). These assays showed that chymotrypsin and elastase cleave pNiXa at the P1-P1 (Thr-Lys) peptide bond near the C-terminus, while trypsin and cathepsin G cleave pNiXa at specific peptide bonds near the N-terminus, within an interesting 26-residue segment, rich in Lys and Gln, that separates the His-cluster of pNiXa from the rest of the molecule. The segment lacks homology to other serpins, but resembles a domain of Xenopus POU3 transcription factor. This study identifies the specific s...Continue Reading

References

Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Jan 1, 1987·Molecular and Cellular Endocrinology·L J Holland, L J Wangh
Dec 1, 1973·Analytical Biochemistry·W Schaffner, C Weissmann
Oct 29, 1993·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·C J Marshall
Jul 1, 1994·Molecular Reproduction and Development·S Grbac-IvankovicF W Sunderman
Mar 1, 1996·The Biochemical Journal·W S ChangR W Carrell
Jan 1, 1996·Analytical Biochemistry·K M HummelW W Lilly
May 1, 1996·Biological Chemistry Hoppe-Seyler·D KepplerB Sordat
May 1, 1997·Nature Structural Biology·D A Lawrence

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Citations

Sep 10, 2005·Analytical and Bioanalytical Chemistry·Tina KnispelEvelin Denkhaus
Dec 4, 2003·Mutation Research·Kazimierz S KasprzakKonstantin Salnikow

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