Limits...
Characterization of Mammalian ADAM2 and Its Absence from Human Sperm.

Choi H, Jin S, Kwon JT, Kim J, Jeong J, Kim J, Jeon S, Park ZY, Jung KJ, Park K, Cho C - PLoS ONE (2016)

Bottom Line: Monkey ADAM2 was found to associate with chaperone proteins in testis.In humans, we identified ADAM2 as a 100-kDa protein in the testis, but failed to detect it in sperm.Taken together, our results provide new information regarding the characteristics of ADAM2 in mammalian species, including humans.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, Korea.

ABSTRACT
The members of the ADAM (a disintegrin and metalloprotease) family are membrane-anchored multi-domain proteins that play prominent roles in male reproduction. ADAM2, which was one of the first identified ADAMs, is the best studied ADAM in reproduction. In the male germ cells of mice, ADAM2 and other ADAMs form complexes that contribute to sperm-sperm adhesion, sperm-egg interactions, and the migration of sperm in the female reproductive tract. Here, we generated specific antibodies against mouse and human ADAM2, and investigated various features of ADAM2 in mice, monkeys and humans. We found that the cytoplasmic domain of ADAM2 might enable the differential association of this protein with other ADAMs in mice. Western blot analysis with the anti-human ADAM2 antibodies showed that ADAM2 is present in the testis and sperm of monkeys. Monkey ADAM2 was found to associate with chaperone proteins in testis. In humans, we identified ADAM2 as a 100-kDa protein in the testis, but failed to detect it in sperm. This is surprising given the results in mice and monkeys, but it is consistent with the failure of ADAM2 identification in the previous proteomic analyses of human sperm. These findings suggest that the reproductive functions of ADAM2 differ between humans and mice. Our protein analysis showed the presence of potential ADAM2 complexes involving yet-unknown proteins in human testis. Taken together, our results provide new information regarding the characteristics of ADAM2 in mammalian species, including humans.

No MeSH data available.


Related in: MedlinePlus

Integrity of human sperm proteins.Western blot analysis was performed using antibodies against ADAM2, PRKACA, PRKARIα, HSPA5, and ADAM7. The TUBA and GAPDH antibodies were used as controls. Experiments were repeated twice. Reduced protein samples were subjected to SDS-PAGE using 10% resolving gels. Abbreviation: NS, non-specific band. Molecular masses are presented on the left.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4920383&req=5

pone.0158321.g007: Integrity of human sperm proteins.Western blot analysis was performed using antibodies against ADAM2, PRKACA, PRKARIα, HSPA5, and ADAM7. The TUBA and GAPDH antibodies were used as controls. Experiments were repeated twice. Reduced protein samples were subjected to SDS-PAGE using 10% resolving gels. Abbreviation: NS, non-specific band. Molecular masses are presented on the left.

Mentions: To ensure the integrity of the human sperm proteins used in the analysis shown in Figs 5 and 6, we carried out Western blot analysis with various antibodies against sperm proteins, including PRKACA, PRKARIα, HSPA5, ADAM7, TUBA, and GAPDH. PRKACA and PRKARIα are the catalytic and regulatory subunits of PKA, respectively, and are known to be present in human sperm [38]. HSPA5 plays an active role in remodeling the sperm surface during capacitation [39]. ADAM7 is synthesized in the epididymis and is known to be secreted and transferred to mature human sperm [40, 41]. All of these marker proteins were detected at their expected molecular sizes (Fig 7), confirming the integrity of the human sperm proteins studied herein.


Characterization of Mammalian ADAM2 and Its Absence from Human Sperm.

Choi H, Jin S, Kwon JT, Kim J, Jeong J, Kim J, Jeon S, Park ZY, Jung KJ, Park K, Cho C - PLoS ONE (2016)

Integrity of human sperm proteins.Western blot analysis was performed using antibodies against ADAM2, PRKACA, PRKARIα, HSPA5, and ADAM7. The TUBA and GAPDH antibodies were used as controls. Experiments were repeated twice. Reduced protein samples were subjected to SDS-PAGE using 10% resolving gels. Abbreviation: NS, non-specific band. Molecular masses are presented on the left.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920383&req=5

pone.0158321.g007: Integrity of human sperm proteins.Western blot analysis was performed using antibodies against ADAM2, PRKACA, PRKARIα, HSPA5, and ADAM7. The TUBA and GAPDH antibodies were used as controls. Experiments were repeated twice. Reduced protein samples were subjected to SDS-PAGE using 10% resolving gels. Abbreviation: NS, non-specific band. Molecular masses are presented on the left.
Mentions: To ensure the integrity of the human sperm proteins used in the analysis shown in Figs 5 and 6, we carried out Western blot analysis with various antibodies against sperm proteins, including PRKACA, PRKARIα, HSPA5, ADAM7, TUBA, and GAPDH. PRKACA and PRKARIα are the catalytic and regulatory subunits of PKA, respectively, and are known to be present in human sperm [38]. HSPA5 plays an active role in remodeling the sperm surface during capacitation [39]. ADAM7 is synthesized in the epididymis and is known to be secreted and transferred to mature human sperm [40, 41]. All of these marker proteins were detected at their expected molecular sizes (Fig 7), confirming the integrity of the human sperm proteins studied herein.

Bottom Line: Monkey ADAM2 was found to associate with chaperone proteins in testis.In humans, we identified ADAM2 as a 100-kDa protein in the testis, but failed to detect it in sperm.Taken together, our results provide new information regarding the characteristics of ADAM2 in mammalian species, including humans.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, Korea.

ABSTRACT
The members of the ADAM (a disintegrin and metalloprotease) family are membrane-anchored multi-domain proteins that play prominent roles in male reproduction. ADAM2, which was one of the first identified ADAMs, is the best studied ADAM in reproduction. In the male germ cells of mice, ADAM2 and other ADAMs form complexes that contribute to sperm-sperm adhesion, sperm-egg interactions, and the migration of sperm in the female reproductive tract. Here, we generated specific antibodies against mouse and human ADAM2, and investigated various features of ADAM2 in mice, monkeys and humans. We found that the cytoplasmic domain of ADAM2 might enable the differential association of this protein with other ADAMs in mice. Western blot analysis with the anti-human ADAM2 antibodies showed that ADAM2 is present in the testis and sperm of monkeys. Monkey ADAM2 was found to associate with chaperone proteins in testis. In humans, we identified ADAM2 as a 100-kDa protein in the testis, but failed to detect it in sperm. This is surprising given the results in mice and monkeys, but it is consistent with the failure of ADAM2 identification in the previous proteomic analyses of human sperm. These findings suggest that the reproductive functions of ADAM2 differ between humans and mice. Our protein analysis showed the presence of potential ADAM2 complexes involving yet-unknown proteins in human testis. Taken together, our results provide new information regarding the characteristics of ADAM2 in mammalian species, including humans.

No MeSH data available.


Related in: MedlinePlus