Inhibition of a novel sperm gelatinase in prawn sperm by the male reproduction-related Kazal-type peptidase inhibitor.
Previously, we have identified and characterized a male reproduction-related kazal-type peptidase inhibitor (MRPINK) gene from the prawn, Macrobrachium rosenbergii. In the present study, MRPINK was discovered to have an inhibitory effect on the gelatinolytic activity of M. rosenbergii sperm and immunofluorescence analysis revealed it bound specifically onto the base of sperm. The proteolytic activity of sperm extracts to vitelline coat components was also detected to be interfered by MRPINK. Furthermore, a novel gelatinase on sperm was found to be specifically inhibited by MRPINK and was named M. rosenbergii sperm gelatinase (MSG). MSG was then isolated and purified by reversed-phase high performance liquid chromatography combining with gelatinolytic assay. By amino-terminal amino acid sequence analysis and molecular cloning, the primary structure of MSG was determined. The data presented in this study provided evidence that MRPINK has an inhibitory effect on the gelatinolytic activity as well as proteolytic activity of prawn sperm and specifically blocks the activity of MSG.
Li
Ye
Y
Institute of Cell Biology and Genetics, College of Life Sciences, Zhejiang University, Zijingang Campus, Hangzhou, Zhejiang, PR China.
Ma
Wen-Ming
WM
Dai
Jie-Qiong
JQ
Feng
Chen-Zhuo
CZ
Yang
Fan
F
Ohira
Tsuyoshi
T
Nagasawa
Hiromichi
H
Yang
Wei-Jun
WJ
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D018093:Gelatinases / Q000235:genetics / Q000378:metabolism*; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D033541:Palaemonidae / Q000378:metabolism*; D011480:Protease Inhibitors / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D013094:Spermatozoa / Q000378:metabolism*; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000378:metabolism*
D017931:DNA Primers; D011480:Protease Inhibitors; D014359:Trypsin Inhibitor, Kazal Pancreatic; D018093:Gelatinases