Cloning and characterization of a novel sperm tail protein, NYD-SP28.
In this study, a gene coding a novel human sperm tail protein named NYD-SP28 was cloned and characterized using a complementary DNA (cDNA) microarray. Its expression was 3.5 times higher in human testis than in fetal testis, and very high in human spermatozoa. The full length of NYD-SP28 cDNA was 1798 bp and encoded a 484-amino-acid protein. Motif analysis revealed that the protein contained a cluster of phosphorylation sites, N-glycosylation sites and N-myristoylation sites. Immunohistochemical analysis of normal human testes showed that NYD-SP28 was expressed in the cytoplasm of spermatogenic cells but not in interstitial cells. The EGFP-NYD-SP28 fusion protein was also localized in the cytoplasm of transfected 7721 cells. In human spermatozoa, NYD-SP28 immunoreactivity was detected in entire sperm tail. Using the two-dimensional (2-D) gel electrophoresis and immunoblotting technique, NYD-SP28 was found to be post-translationally modified during sperm capacitation. In conclusion, these results suggest that NYD-SP28 is a new human sperm tail protein and might play an important role during sperm capacitation.
Zheng
Ying
Y
Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 10029, PR China.
Zhang
Jingjing
J
Wang
Lei
L
Zhou
Zuomin
Z
Xu
Min
M
Li
Jianmin
J
Sha
Jia-Hao
JH
D000328:Adult; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D002874:Chromosome Mapping; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary; D005091:Exons; D005333:Fetus; D020869:Gene Expression Profiling; D006023:Glycoproteins; D006801:Humans; D007150:Immunohistochemistry; D007526:Isoelectric Point; D008113:Liver Neoplasms / Q000473:pathology; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012661:Semen / Q000166:cytology; D017386:Sequence Homology, Amino Acid; D013082:Sperm Tail / Q000737:chemistry* / Q000378:metabolism; D013094:Spermatozoa / Q000166:cytology; D013737:Testis / Q000737:chemistry / Q000166:cytology; D014162:Transfection
C516344:CCDC65 protein, human; D018076:DNA, Complementary; D006023:Glycoproteins; D011506:Proteins; D011993:Recombinant Fusion Proteins