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Identification of Goat Sperm Ecto-Cyclic AMP Independent Protein Kinase Substrate Localized on Sperm Outer Surface

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Title Identification of Goat Sperm Ecto-Cyclic AMP
Independent Protein Kinase Substrate Localized
on Sperm Outer Surface
 
Creator Maiti, Arunima
Mishra, K.P
Majumder, Gopal Chandra
 
Subject Cell Biology & Physiology
 
Description We have demonstrated the location of a cyclic AMP independent serine/threonine protein kinase
(ecto-CIK) on the outer surface of mature goat spermatozoa. We purified and characterized the major physiological
protein substrate (MPS) of ecto-CIK. 32P-labeled membrane proteins phosphorylated by endogenous ecto-CIK of intact
cauda-epididymal spermatozoa was solubilized with 1% Triton X-100 and then fractionated by following several
chromatographic techniques like Sephacryl S-300 molecular sieve chromatography, DEAE–cellulose ion-exchange
chromatography and chromatofocussing. The MPS of ecto-CIK has been purified to apparent homogeneity and it was
found to be a monomeric protein of 100 kDa. Three isoforms of MPS have been found with pI of 6.37, 6.05, and 5.14 and
all these isoforms served as the specific substrate of ecto-CIK. The ecto-kinase has nearly 30 times greater affinity forMPSas
compared to casein the most potent exogenous protein substrate. Addition of MPS (pI 5.14) antibody caused head-to-head
sperm agglutination. The Fv/Fab fragment of anti-MPS caused significant inhibition of sperm motility. The data show that
MPS is an ecto-protein localized on the sperm head. MPS may thus play an important role for the regulation of sperm–egg
interaction. J. Cell. Biochem. 92: 164–177, 2004. � 2004 Wiley-Liss, Inc
 
Publisher John Wiley & Sons
 
Date 2004
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/538/1/JOURNAL_OF_CELLULAR_BIOCHEMISTRY%2C92(_1)%2C164%2D177_[54].pdf
Maiti, Arunima and Mishra, K.P and Majumder, Gopal Chandra (2004) Identification of Goat Sperm Ecto-Cyclic AMP Independent Protein Kinase Substrate Localized on Sperm Outer Surface. Journal of Cellular Biochemistry, 92 (1). pp. 164-177. ISSN 0730-2312
 
Relation http://dx.doi.org/10.1002/jcb.20060
http://www.eprints.iicb.res.in/538/