Certain Strongylocentrotus purpuratus sperm mitochondrial proteins co-purify with low density detergent-insoluble membranes and are PKA or PKC-substrates possibly involved in sperm motility regulation
Por:
Loza-Huerta, A, Vera-Estrella, R, Darszon, A, Beltran, C
Publicada:
1 nov 2013
Resumen:
Background: Sea urchin sperm motility is regulated by Speract, a
sperm-activating peptide (SAP) secreted from the outer egg coat. Upon
binding to its receptor in the sperm flagellum, Speract induces a series
of ionic and metabolic changes in Strongylocentrotus purpuratus
spermatozoa that regulate their motility. Among these events, protein
phosphorylation is one of the most relevant and evidence indicates that
some proteins of the Speract signaling cascade localize in low density
detergent-insoluble membranes (LD-DIM).
Methods: LD-DIM-derived proteins from immotile, motile or
Speract-stimulated S. purpuratus sperm were resolved in 2-D gels and the
PKA and PKC substrates detected with specific antibodies were identified
by LCMS/MS.
Results: Differential PICA and PKC substrate phosphorylation levels
among the LD-DIM isolated from sperm in different motility conditions
were foiind and identified by mass spectrometry as: ATP synthase,
creatine kinase, NADH dehydrogenase (ubiquinone) flavoprotein 2,
succinyl-CoA ligase and the voltage-dependent anion channel 2 (VDAC2),
which are mitochondrial proteins, as well as, the cAMP-dependent protein
kinase type II regulatory (PICA RII) subunit, Tubulin beta chain and
Actin Cy I changed their phosphorylation state. Conclusions: Some
mitochondrial proteins regulated by PICA or PKC may influence sea urchin
sperm motility.
General significance: The fact that a high percentage (66%) of the PICA
or PKC substrates identified in LD-DIM are mitochondrial proteins
suggests that the phosphorylation of these proteins modulates sea urchin
sperm motility via Speract stimulation by providing sufficient energy to
sperm physiology. Those mitochondrial proteins are indeed PICA- or
PKC-substrates in the sea urchin spermatozoa. (C) 2013 Elsevier B.V. All
rights reserved.
Filiaciones:
Loza-Huerta, A:
Univ Nacl Autonoma Mexico, Dept Genet Desarrollo & Fisiol Mol, Cuernavaca 62210, Morelos, Mexico
Vera-Estrella, R:
Univ Nacl Autonoma Mexico, Inst Biotecnol, Cuernavaca 62210, Morelos, Mexico
Darszon, A:
Univ Nacl Autonoma Mexico, Dept Genet Desarrollo & Fisiol Mol, Cuernavaca 62210, Morelos, Mexico
Beltran, C:
Univ Nacl Autonoma Mexico, Dept Genet Desarrollo & Fisiol Mol, Cuernavaca 62210, Morelos, Mexico
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