Molecular mechanism of epididymal protease inhibitor modulating the liquefaction of human semen / 亚洲男科学杂志(英文版)
Asian Journal of Andrology
; (6): 770-775, 2008.
Article
em En
| WPRIM
| ID: wpr-359911
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>AIM</b>To study the molecular mechanism of epididymal protease inhibitor (Eppin) modulating the process of prostate specific antigen (PSA) digesting semenogelin (Sg).</p><p><b>METHODS</b>Human Sg cDNA (nucleotides 82-849) and Eppin cDNA (nucleotides 70-723) were generated by polymerase chain reaction (PCR) and cloned into pET-100D/TOPO. Recombinant Eppin and Sg (rEppin and rSg) were produced by BL21 (DE3). The association of Eppin with Sg was studied by far-western immunoblot and radioautography. In vitro the digestion of rSg by PSA in the presence or absence of rEppin was studied. The effect of anti-Q20E (N-terminal) and C-terminal of Eppin on Eppin-Sg binding was monitored.</p><p><b>RESULTS</b>Eppin binds Sg on the surface of human spermatozoa with the C-terminal of Eppin (amino acids 75-133). rSg was digested with PSA and many low molecular weight fragments were produced. When rEppin is bound to rSg, then digested by PSA, incomplete digestion and a 15-kDa fragment results. Antibody binding to the N-terminal of rEppin did not affect rSg digestion. Addition of antibodies to the C-terminal of rEppin inhibited the modulating effect of rEppin.</p><p><b>CONCLUSION</b>Eppin protects a 15-kDa fragment of rSg from hydrolysis by PSA.</p>
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Índice:
WPRIM
Assunto principal:
Farmacologia
/
Sêmen
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Espermatozoides
/
Autorradiografia
/
Proteínas Recombinantes
/
Antígeno Prostático Específico
/
Biologia Celular
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Proteínas Secretadas pela Vesícula Seminal
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Alergia e Imunologia
/
Proteínas Secretadas Inibidoras de Proteinases
Limite:
Animals
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Humans
/
Male
Idioma:
En
Revista:
Asian Journal of Andrology
Ano de publicação:
2008
Tipo de documento:
Article