Cysteine-rich secretory protein 2 binds to mitogen-activated protein kinase kinase kinase 11 in mouse sperm.
Cysteine-rich secretory protein (CRISP) 2 (previously TPX1) is a testis-enriched member of the CRISP family, and has been localized to both the sperm acrosome and tail. Like all members of the mammalian CRISP family, its expression pattern is strongly suggestive of a role in male fertility, but functional support for this hypothesis remains limited. In order to determine the biochemical pathways within which CRISP2 is a component, the putative mature form of CRISP2 was used as bait in a yeast two-hybrid screen of a mouse testis expression library. One of the most frequently identified interacting partners was mitogen-activated protein kinase kinase kinase 11 (MAP3K11). Sequencing and deletion experiments showed that the carboxyl-most 20 amino acids of MAP3K11 interacted with the CRISP domain of CRISP2. This interaction was confirmed using pull-down experiments and the cellular context was supported by the localization of CRISP2 and MAP3K11 to the acrosome of the developing spermatids and epididymal spermatozoa. Interestingly, mouse epididymal sperm contained an approximately 60-kDa variant of MAP3K11, which may have been a result of proteolytic cleavage of the longer 93-kDa form seen in many tissues. These data raise the possibility that CRISP2 is a MAP3K11-modifying protein or, alternatively, that MAP3K11 acts to phosphorylate CRISP2 during acrosome development.
Gibbs
Gerard M
GM
Monash Institute of Medical Research, Monash University, Melbourne, Victoria 3168, Australia.
Bianco
Deborah M
DM
Jamsai
Duangporn
D
Herlihy
Amy
A
Ristevski
Sika
S
Aitken
R John
RJ
Kretser
David M de
DM
O'Bryan
Moira K
MK
D000818:Animals; D015815:Cell Adhesion Molecules; D004822:Epididymis / Q000166:cytology; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D020930:MAP Kinase Kinase Kinases / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*; D013737:Testis / Q000166:cytology; D000097767:Mitogen-Activated Protein Kinase Kinase Kinase 11
D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D020930:MAP Kinase Kinase Kinases; D008565:Membrane Proteins; D000097767:Mitogen-Activated Protein Kinase Kinase Kinase 11; C483816:Crisp2 protein, mouse