scholarly journals Participation of the Human Sperm Proteasome in the Capacitation Process and Its Regulation by Protein Kinase A and Tyrosine Kinase1

2009 ◽  
Vol 80 (5) ◽  
pp. 1026-1035 ◽  
Author(s):  
Milene Kong ◽  
Emilce S. Diaz ◽  
Patricio Morales
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Human Sperm ◽  
Kinase A

scholarly journals The activation of the chymotrypsin-like activity of the proteasome is regulated by soluble adenyl cyclase/cAMP/protein kinase A pathway and required for human sperm capacitation

2019 ◽  
Vol 25 (10) ◽  
pp. 587-600 ◽  
Author(s):  
Héctor Zapata-Carmona ◽  
Lina Barón ◽  
Lidia M Zuñiga ◽  
Emilce Silvina Díaz ◽  
Milene Kong ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Current Knowledge ◽  
Human Sperm ◽  
Adenyl Cyclase ◽  
Sperm Capacitation ◽  
Time Treatment ◽  
Proteasome Subunits ◽  
Pka Pathway ◽  
Kinase A

Abstract One of the first events of mammalian sperm capacitation is the activation of the soluble adenyl cyclase/cAMP/protein kinase A (SACY/cAMP/PKA) pathway. Here, we evaluated whether the increase in PKA activity at the onset of human sperm capacitation is responsible for the activation of the sperm proteasome and whether this activation is required for capacitation progress. Viable human sperm were incubated with inhibitors of the SACY/cAMP/PKA pathway. The chymotrypsin-like activity of the sperm proteasome was evaluated using a fluorogenic substrate. Sperm capacitation status was evaluated using the chlortetracycline assay and tyrosine phosphorylation. To determine whether proteasomal subunits were phosphorylated by PKA, the proteasome was immunoprecipitated and tested on a western blot using an antibody against phosphorylated PKA substrates. Immunofluorescence microscopy analysis and co-immunoprecipitation (IPP) were used to investigate an association between the catalytic subunit alpha of PKA (PKA-Cα) and the proteasome. The chymotrypsin-like activity of the sperm proteasome significantly increased after 5 min of capacitation (P < 0.001) and remained high for the remaining incubation time. Treatment with H89, KT5720 or KH7 significantly decreased the chymotrypsin-like activity of the proteasome (P < 0.001). IPP experiments indicated that PKA inhibition significantly modified phosphorylation of proteasome subunits. In addition, PKA-Cα colocalized with the proteasome in the equatorial segment and in the connecting piece, and co-immunoprecipitated with the proteasome. This is the first demonstration of sperm proteasome activity being directly regulated by SACY/PKA-Cα. This novel discovery extends our current knowledge of sperm physiology and may be used to manage sperm capacitation during assisted reproductive technology procedures.

Interaction Between Protein Kinase A (PKA) and Protein Phosphatase PP2A During Human Sperm Capacitation.

2012 ◽  
Vol 87 (Suppl_1) ◽  
pp. 448-448
Author(s):  
Patricio J. Morales ◽  
Kely Ordenes ◽  
Lidia Zuñiga ◽  
Emilce S. Diaz
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Protein Phosphatase ◽  
Human Sperm ◽  
Sperm Capacitation ◽  
Kinase A

Protein Kinase A modulation by nitric oxide during human sperm capacitation

2018 ◽  
Author(s):  
Florentin-Daniel Staicu ◽  
CARMEN MATAS PARRA ◽  
Juan Carlos Martínez Soto
Keyword(s):  
Nitric Oxide ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Human Sperm ◽  
Sperm Capacitation ◽  
Kinase A

scholarly journals Protein kinase A inhibition induces EPAC-dependent acrosomal exocytosis in human sperm

2019 ◽  
Vol 21 (4) ◽  
pp. 337 ◽  
Author(s):  
Haim Breitbart ◽  
Diana Itzhakov ◽  
Yeshayahu Nitzan
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Human Sperm ◽  
Acrosomal Exocytosis ◽  
Kinase A

scholarly journals Involvement of tyrosine kinase and cAMP-dependent kinase cross-talk in the regulation of human sperm motility

Reproduction ◽  
2003 ◽  
pp. 183-195 ◽  
Author(s):  
M Bajpai ◽  
GF Doncel
Keyword(s):  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Protein Kinase A ◽  
Tyrosine Phosphorylation ◽  
Human Sperm ◽  
Human Spermatozoa ◽  
Phosphorylated Proteins ◽  
Motion Parameters ◽  
Sperm Motion ◽  
Kinase A

Tyrosine phosphorylation and its upregulation by cAMP have been associated with capacitation and motility changes of spermatozoa. In the present study, washed spermatozoa were incubated for 6 h in protein-supplemented complete medium with or without kinase inhibitors to verify whether upstream activation of protein kinase A is indispensable for tyrosine phosphorylation and motility changes to occur in capacitating human spermatozoa. H89, a specific protein kinase A inhibitor, significantly inhibited the activity of sperm protein kinase A. However, this inhibition did not alter capacitation-related tyrosine kinase activation. Tyrosine phosphorylated proteins, motion parameters and the incidence of phosphotyrosine-immunoreactive spermatozoa were decreased only slightly. Conversely, genistein, a tyrosine kinase inhibitor which inhibited sperm tyrosine kinase but not protein kinase A, significantly reduced all the parameters studied. Spermatozoa incubated with cAMP and pentoxifylline showed a rapid enhancement of tyrosine phosphorylation and some of the sperm motion parameters, particularly hyperactivation. Inclusion of H89 reduced cAMP stimulation of tyrosine kinase, and tyrosine phosphorylation and motion parameters were reduced almost to basal values. Treatment with genistein reduced tyrosine kinase activity, especially in the soluble fraction of sperm extracts. A decrease in tyrosine phosphorylation of soluble proteins, 105, 81, 55 and 48 kDa, correlated with a significant reduction in sperm motion parameters. Hyperactivation was reduced by tenfold. Tyrosine phosphorylated proteins in the insoluble fraction and the incidence of tyrosine phosphorylated-positive spermatozoa were not reduced markedly. Upstream protein kinase A activation may be a facilitatory rather than an indispensable step in the capacitation-induced tyrosine phosphorylation mediating motility changes in human spermatozoa. Triton-x100 soluble tyrosine phosphorylated proteins, more than their insoluble counterparts, appear to be involved in the modulation of human sperm motion characteristics.

scholarly journals Protein Kinase A (PRKA) Activity Is Regulated by the Proteasome at the Onset of Human Sperm Capacitation

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3501
Author(s):  
Héctor Zapata-Carmona ◽  
Lina Barón ◽  
Milene Kong ◽  
Patricio Morales
Keyword(s):  
Protein Kinase ◽  
Western Blot ◽  
Protein Kinase A ◽  
Human Sperm ◽  
Sperm Capacitation ◽  
Phosphorylation Pattern ◽  
Presence And Absence ◽  
Kinase A

The proteasome increases its activity at the onset of sperm capacitation due to the action of the SACY/PRKACA pathway; this increase is required for capacitation to progress. PRKA activity also increases and remains high during capacitation. However, intracellular levels of cAMP decrease in this process. Our goal was to evaluate the role of the proteasome in regulating PRKA activity once capacitation has started. Viable human sperm were incubated in the presence and absence of epoxomicin or with 0.1% DMSO. The activity of PRKA; the phosphorylation pattern of PRKA substrates (pPRKAs); and the expression of PRKAR1, PRKAR2, and AKAP3 were evaluated by Western blot. The localization of pPRKAs, PRKAR1, PRKAR2, and AKAP3 was evaluated by immunofluorescence. Treatment with epoxomicin changed the localization and phosphorylation pattern and decreased the percentage of pPRKAs-positive sperm. PRKA activity significantly increased at 1 min of capacitation and remained high throughout the incubation. However, epoxomicin treatment significantly decreased PRKA activity after 30 min. In addition, PRKAR1 and AKAP3 were degraded by the proteasome but with a different temporal kinetic. Our results suggest that PRKAR1 is the target of PRKA regulation by the proteasome.

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