Human ribonuclease 9, a member of ribonuclease A superfamily, specifically expressed in epididymis, is a novel sperm-binding protein.
- Author:
Gui-Zhi CHENG
1
;
Jian-Yuan LI
;
Fang LI
;
Hai-Yan WANG
;
Guang-Xia SHI
Author Information
- Publication Type:Journal Article
- MeSH: Adult; Amino Acid Sequence; Anti-Infective Agents; metabolism; Blotting, Western; Cloning, Molecular; Enzyme-Linked Immunosorbent Assay; Epididymis; enzymology; Escherichia coli; enzymology; Genetic Vectors; Humans; Male; Molecular Sequence Data; Recombinant Fusion Proteins; chemistry; metabolism; Ribonuclease, Pancreatic; metabolism; Ribonucleases; chemistry; metabolism; Seminal Plasma Proteins; chemistry; metabolism; Spermatozoa; metabolism; Testis; enzymology; Young Adult
- From: Asian Journal of Andrology 2009;11(2):240-251
- CountryChina
- Language:English
- Abstract: To explore the functions of human ribonuclease 9 (RNase 9), we constructed a mammalian fusion expression vector pcDNA-hRNase9, prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences. According to the determined mature protein, recombinant human RNase 9 was prepared in E. coli. Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected, and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay. The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA, but exhibited antibacterial activity, in a concentration/time dependent manner, against E. coli. Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis, but not present in other tissues examined, and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa. These results suggest that human RNase 9 may play roles in host defense of male reproductive tract.
