Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
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Query: UNIPROT:Q9UIJ5 (
Rec
)
58,342
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Proacrosin is a multifunctional protein present in the sperm acrosome. This study characterizes the expression of human proacrosin in bacteria and assesses zona pellucida binding activity. The cDNA encoding human proacrosin was subcloned in pGEX-3X and pET-22b vectors. In the pGEX system, expression of the full-length fusion protein was not detected. In the pET system, an expression product with an apparent molecular size similar to that expected for the proenzyme (Rec-40, 42-44 kDa) was recognized by a monoclonal antibody to human
acrosin
, AcrC5F10. A 32-34-kDa protein (Rec-30), not recognized by AcrC5F10 on Western blots, was the major expression product. Proteins of 21 (Rec-20) and 18 (Rec-10) kDa were recovered as insoluble expression products as were
Rec
-40 and
Rec
-30, and truncated products from the C terminus were detected in the soluble fraction.
Rec
-40 and
Rec
-30 coexisted at any culture time tested. Immune serum raised against
Rec
-30 (AntiRec-30) stained the acrosomal region of permeabilized human spermatozoa and recognized the recombinant proteins and proacrosin from human sperm extracts. Amino acid sequence analysis indicated that
Rec
-30,
Rec
-20, and
Rec
-10 are N-terminal fragments of proacrosin. The recombinant proteins
Rec
-40, -30, -20, and -10 were found to interact with homologous (125)I-zona pellucida glycoproteins.
...
PMID:Expression of human proacrosin in Escherichia coli and binding to zona pellucida. 1068 1
Recently, the endocannabinoid (EC) system and the presence of CB1 receptor (CB1-R), have been identified in human sperm. However, the effects of EC receptor ligands such as anandamide (N-arachidonoylethanolamine) and the role of EC system in male fertility is still largely unexplored. In the present study, we investigated the ultrastructural compartmentalization of CB1-R and analyzed the effects of its stimulation by using a stable analog of anandamide, 2-methylarachidonyl-2'-fluoro-ethylamide (MET-F-AEA). We focused particularly on sperm survival and
acrosin
activity. The study of human sperm anatomy by transmission electron microscopy with immunogold analysis revealed the location of the CB1-R prevalently in the sperm membranes of the head and interestingly on the mitochondria. The effect of different concentrations of MET-F-AEA from 100 nM to 1 microM evidenced a significant decrease of sperm survival. Interestingly, we analyzed this negative effect at molecular level, testing the EC action on different known sperm survival targets. MET-F-AEA-treatment decreased both pBCL2 and pAkt, two prosurvival proteins, and increased pPTEN expression which is the main regulator of the PI3K/Akt pathway. Moreover, a biphasic effect was observed with increasing MET-F-AEA concentrations on the
acrosin
activity. The blockage of the CB1-R by using its selective antagonist SR141716 (rimonabant) induced an opposite action on sperm survival supporting a role for this receptor in the biology of the male gamete.
Anat
Rec
(Hoboken) 2010 Feb
PMID:Human sperm anatomy: ultrastructural localization of the cannabinoid1 receptor and a potential role of anandamide in sperm survival and acrosome reaction. 1993 10