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Cathepsin A protein from the accessory sex gland of the Chinese mitten crab (Eriocheir sinensis) plays a key role in spermatophore digestion.

Abstract
Accessory sex gland (ASG) secretory proteins of the Chinese mitten crab (Eriocheir sinensis) can effectively digest the spermatophore wall. In order to identify which proteins participate in spermatophore wall digestion, a 50-kDa protein secreted from the ASG was purified to homogeneity by a series of isolation steps, including ammonium sulfate fractionation, Sephadex G-25 S gel-filtration, ion exchange chromatography on a DEAE-Sephacel column and Sephacryl S-200 gel-filtration. The purified protein was effective in spermatophore wall rupture, and the subsequent HPLC-ESI-MS/MS shotgun analysis showed the digestive protein to be cathepsin A (cathA). This finding was also confirmed by Western blot analysis and a cathA inhibitor digestion experiment. ELISA analysis showed that cathA enzymatic activity from ASG secretions increased during its purification process. Furthermore, enzymatic activity was significantly higher in the mating period of E. sinensis parallel to the latest developmental stage of the gland. Moreover, analysis from a cathA inhibitor that inhibits spermatophore wall digestion showed that cathA is the main enzyme involved. Hence, we first report the characterization of cathA from the ASG, which might play a key role in digesting the spermatophore wall of E. sinensis.
AuthorsJuan Wang, Di-An Fang, Yang Wang, Yuan-Li Wang, Lin Cheng, Lin He, Qun Wang
JournalJournal of insect physiology (J Insect Physiol) Vol. 59 Issue 9 Pg. 953-60 (Sep 2013) ISSN: 1879-1611 [Electronic] England
PMID23871752 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2013 Elsevier Ltd. All rights reserved.
Chemical References
  • Cathepsin A
Topics
  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Cathepsin A (chemistry, metabolism)
  • Decapoda (Crustacea) (metabolism)
  • Genitalia, Male (metabolism)
  • Male
  • Molecular Sequence Data
  • Spermatogonia (metabolism)

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