SPATA2
Spermatogenezi-pridruženi protein 2 jest protein koji je kod ljudi kodiran genom SPATA2.[5]
Kloniranje i ekspresija
Sekvenciranjem klonova dobijenih iz frakcionirane biblioteke cDNK mozga, Nagase et al. (1998) klonirali su SPATA2, koji su označili KIAA0757. Izvedeni protein sadrži 520 aminokiselina. RT-PCR ELISA otkrila je nisku do umjerenu ekspresiju u svim testiranim tkivima, uključujući testise. Graziotto et al. (1999) izolirali su fragment SPATA2, koji su označili kao PD1, i pojačali su cDNK pune dužine, pomoću 5-obazne RACE biblioteke cDNK testisa. Izvedeni protein s 520 aminokiselina ima izračunatu molekulsku masu od 58,4 kD i dijeli oko 33% identiteta sa RBM, proteinom koji veže RNK i koji je uključen u spermatogenezu. SPATA2 je visoko hidrofilni. Nema signalni peptid, bogat je serinom i sadrži 17 ostataka cisteina koji se pretežno nalaze u C-terminalnom domenu. SPATA2 također ima nekoliko potencijalnih mjesta fosforilacije, ali nema mjesta N-glikozilacije.
Northern blot analizom otkriven je obilni prijepis od 2,7 kb i rjeđa trankripcija od 4,0 kb, s najvećom ekspresijom u testisima. Slabija ekspresija otkrivena je u slezeni, zatim timusu i prostati. SPATA2 normalno nije otkriven u leukocitima maternice, tankog i debelog crijeva ili periferne krvi. Western blot analizom ekstrakata ćelija testisa, otkriven je protein s prividnom molekulskm masom od 60 kD, kao i manje obilan protein od 46 do 48 kD. Ekspresija oba proteina, posebno manjeg , smanjena je u nekoliko testikulopatija. Imunohistohemijska analiza otkrila je SPATA2 u citoplazmi Sertolijevih ćelija. SPATA2 nije otkriven u spermatogenim ćelijama i Leydigovim ćelijama, a jedra su bila negativna u svim tipovima ćelija.[6][7]
Aminokiselinska sekvenca
- Simboli
C: Cistein
D: Asparaginska kiselina
E: Glutaminska kiselina
F: Fenilalanin
G: Glicin
H: Histidin
I: Izoleucin
K: Lizin
L: Leucin
M: Metionin
N: Asparagin
P: Prolin
Q: Glutamin
R: Arginin
S: Serin
T: Treonin
V: Valin
W: Triptofan
Y: Tirozin
| 10 | 20 | 30 | 40 | 50 | ||||
|---|---|---|---|---|---|---|---|---|
| MGKPSSMDTK | FKDDLFRKYV | QFHESKVDTT | TSRQRPGSDE | CLRVAASTLL | ||||
| SLHKVDPFYR | FRLIQFYEVV | ESSLRSLSSS | SLRALHGAFS | MLETVGINLF | ||||
| LYPWKKEFRS | IKTYTGPFVY | YVKSTLLEED | IRAILSCMGY | TPELGTAYKL | ||||
| RELVETLQVK | MVSFELFLAK | VECEQMLEIH | SQVKDKGYSE | LDIVSERKSS | ||||
| AEDVRGCSDA | LRRRAEGREH | LTASMSRVAL | QKSASERAAK | DYYKPRVTKP | ||||
| SRSVDAYDSY | WESRKPPLKA | SLSLRKEPVA | TDVGDDLKDE | IIRPSPSLLT | ||||
| MASSPHGSPD | VLPPASPSNG | PALLRGTYFS | TQDDVDLYTD | SEPRATYRRQ | ||||
| DALRPDVWLL | RNDAHSLYHK | RSPPAKESAL | SKCQSCGLSC | SSSLCQRCDS | ||||
| LLTCPPASKP | SAFPSKASTH | DSLAHGASLR | EKYPGQTQGL | DRLPHLHSKS | ||||
| KPSTTPTSRC | GFCNRPGATN | TCTQCSKVSC | DACLSAYHYD | PCYKKSELHK | ||||
| FMPNNQLNYK | STQLSHLVYR |
Reference
- ^ a b c GRCh38: Ensembl release 89: ENSG00000158480 - Ensembl, maj 2017
- ^ a b c GRCm38: Ensembl release 89: ENSMUSG00000047030 - Ensembl, maj 2017
- ^ "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
- ^ "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
- ^ "Entrez Gene: SPATA2 spermatogenesis associated 2".
- ^ Graziotto, R., Foresta, C., Scannapieco, P., Zeilante, P., Russo, A., Negro, A., Salmaso, R., Onisto, M. cDNA cloning and characterization of PD1: a novel human testicular protein with different expressions in various testiculopathies. Exp. Res. Commun. 248: 620-626, 1999.
- ^ Nagase, T., Ishikawa, K., Suyama, M., Kikuno, R., Miyajima, N., Tanaka, A., Kotani, H., Nomura, N., Ohara, O. Prediction of the coding sequences of unidentified human genes. XI. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res. 5: 277-286, 1998. [PubMed]: 9872452
Dopunska literatura
- Bonaldo MF, Lennon G, Soares MB (1997). "Normalization and subtraction: two approaches to facilitate gene discovery". Genome Res. 6 (9): 791–806. doi:10.1101/gr.6.9.791. PMID 8889548.
- Nagase T, Ishikawa K, Suyama M, et al. (1999). "Prediction of the coding sequences of unidentified human genes. XI. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro". DNA Res. 5 (5): 277–86. doi:10.1093/dnares/5.5.277. PMID 9872452.
- Graziotto R, Foresta C, Scannapieco P, et al. (1999). "cDNA cloning and characterization of PD1: a novel human testicular protein with different expressions in various testiculopathies". Exp. Cell Res. 248 (2): 620–6. doi:10.1006/excr.1999.4449. PMID 10222154.
- Deloukas P, Matthews LH, Ashurst J, et al. (2002). "The DNA sequence and comparative analysis of human chromosome 20". Nature. 414 (6866): 865–71. doi:10.1038/414865a. PMID 11780052.
- Strausberg RL, Feingold EA, Grouse LH, et al. (2003). "Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences". Proc. Natl. Acad. Sci. U.S.A. 99 (26): 16899–903. doi:10.1073/pnas.242603899. PMC 139241. PMID 12477932.
- Liliana Slongo M, Zotti L, Onisto M (2003). "Cloning and characterization of the promoter region of human spata2 (spermatogenesis-associated protein 2) gene". Biochim. Biophys. Acta. 1625 (2): 192–6. doi:10.1016/s0167-4781(02)00594-8. PMID 12531478.
- Ota T, Suzuki Y, Nishikawa T, et al. (2004). "Complete sequencing and characterization of 21,243 full-length human cDNAs". Nat. Genet. 36 (1): 40–5. doi:10.1038/ng1285. PMID 14702039.
- Gerhard DS, Wagner L, Feingold EA, et al. (2004). "The Status, Quality, and Expansion of the NIH Full-Length cDNA Project: The Mammalian Gene Collection (MGC)". Genome Res. 14 (10B): 2121–7. doi:10.1101/gr.2596504. PMC 528928. PMID 15489334.
- Rual JF, Venkatesan K, Hao T, et al. (2005). "Towards a proteome-scale map of the human protein-protein interaction network". Nature. 437 (7062): 1173–8. doi:10.1038/nature04209. PMID 16189514. S2CID 4427026.
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