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Sugii Manabu

Affiliate Master Yamaguchi University

Purification of GVBD-inducing protein from the ciliate tetrahymena thermophila

The journal of eukaryotic microbiology Volume 48 Issue 4 Page 414-424
published_at 2001-07
Title
Purification of GVBD-inducing protein from the ciliate tetrahymena thermophila
Creators Sugii Manabu
Creators Fujishima Masahiro
Creator Keywords
cdc2 homologue cell cycle M-phase promoting factor PSTAIR antibody p 13^<sue1>-beads Xenopus oocytes
Germinal-vesicle-breakdown (GVBD) was induced if a 132,000-g supernatant of Tetrahymena thermophila homogenates was injected into Xenopus oocytes. Using this induction of GVBD as a bioassay system, a GVBD-inducing substance was purified from the Tetrahymena by ultra-filtration, liquid chromatography, and electroelution from a band on native-PAGE gel. Proteins eluted from the single band on the native-PAGE gel induced GVBD in the absence of oocyte protein synthesis. This band resolved into two bands on SDS-PAGE: 60 and 1 12 kDa. The 60 kDa protein was the active fraction inducing GVBD. Immunoprecipitation of the 60 kDa protein prevented the GVBD-inducing activity, supporting the conclusion that the 60 kDa protein is the GVBD-inducing substance. An immunoblot with anti-60 kDa monoclonal antibody and PSTAIR antibody showed that p 13^<sue1>-beads could remove cdc2 homologues from T. thermophila supernatant but could not remove the GVBD-inducing activity. The 60-kDa protein appeared at the same time as micronuclear division and disappeared at the beginning of the macronuclear division during synchronous cell division. The cyclic appearance of the 60-kDa protein in the T. thermophila cell cycle suggests that this protein has a cell cycle function.
Languages eng
Resource Type journal article
Publishers Blackwell
Date Issued 2001-07
File Version Not Applicable (or Unknown)
Access Rights metadata only access
Relations
[ISSN]1066-5234
[NCID]AA10895978
info:doi/10.1111/j.1550-7408.2001.tb00174.x
[isVersionOf] [URI]http://www.blackwell-synergy.com/loi/jeu
Schools 大学情報機構 大学院理工学研究科(理学)