Click here to close Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly. We suggest using a current version of Chrome, FireFox, or Safari.
XB-ART-14294
Comp Biochem Physiol B Biochem Mol Biol March 1, 1998; 119 (3): 571-6.
Show Gene links Show Anatomy links

Purification and properties of embryonic cysteine proteinase which participates in yolk-lysis of Xenopus laevis.

Yoshizaki N , Moriyama A , Yonezawa S .


Abstract
The study reported here aimed to purify a cysteine proteinase from neurula embryos of Xenopus laevis, since this enzyme was thought to be involved in yolk-lysis in developing embryos. The purification procedure consisted of fractionation of an embryonic extract by means of 30-90% ammonium sulfate, chromatography on diethylaminoethyl cellulose and carboxymethyl cellulose, gel filtration on Sephadex G-75 and affinity chromatography on concanavalin A-agarose. The purified enzyme had a molecular weight of 30 kDa according to both SDS-PAGE and Sephadex G-75 gel-filtration and an optimum pH of 5.5, and it preferentially cleaved the synthetic substrate, Z-Phe-Arg-MCA. Its activity was inhibited by Z-Phe-Phe-CHN2, a specific cathepsin L inhibitor, as well as by leupeptin and E-64. The NH2-terminal amino acid sequence of the enzyme was similar to that of chicken cathepsin B. These characteristics indicate that the purified enzyme is a member of the cysteine proteinase family. The antibody raised against the purified enzyme specifically stained a 30 kDa protein of neurula embryo extracts on immunoblot tests. The enzyme effectively digested Xenopus yolk proteins when the NaCl concentration in test solutions was 0.2 M. It was also confirmed that cysteine proteinase inhibitors inhibited yolk-lysis by the enzyme.

PubMed ID: 9734341
Article link:


Species referenced: Xenopus laevis
Genes referenced: chn1 chn2 ctsb ctsl