Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility

Srinivasan Vijayaraghavan, Said A. Goueli, Michael Davey, Daniel Carr

Research output: Contribution to journalArticle

196 Citations (Scopus)

Abstract

Cyclic AMP-dependent protein kinase (PKA) is anchored at specific subcellular sites through the interaction of the regulatory subunit (R) with protein kinase A-anchoring proteins (AKAPs) via an amphipathic helix binding motif. Synthetic peptides containing this amphipathic helix domain competitively disrupt PKA binding to AKAPs and cause a loss of PKA modulation of cellular responses. In this report we use S-Ht31, a cell-permeant anchoring inhibitor peptide, to study the role of PKA anchoring in sperm. Our analysis of three species of mammalian sperm detected three isoforms of PKA (RIIα, RIIβ, and RIβ) and one 110-kDa AKAP. The addition of S-Ht31 to bovine caudal epididymal sperm inhibits motility in a time- and concentration-dependent manner. A control peptide, S-Ht31-P, identical to S- Ht31 except for a proline for isoleucine substitution to prevent amphipathic helix formation, had no effect on motility. The inhibition of motility by S- Ht31 is reversible but only if calcium is present in the suspension buffer, suggesting a role for PKA anchoring in regulating cellular calcium homeostasis. Surprisingly, inhibition of PKA catalytic activity had little effect on basal motility or motility stimulated by agents previously thought to work via PKA activation. These data suggest that the interaction of the regulatory subunit of PKA with sperm AKAPs, independent of PKA catalytic activity, is a key regulator of sperm motility and that disruption of this interaction using cell-permeable anchoring inhibitor peptides may form the basis of a sperm-targeted contraceptive.

Original languageEnglish (US)
Pages (from-to)4747-4752
Number of pages6
JournalJournal of Biological Chemistry
Volume272
Issue number8
DOIs
StatePublished - Feb 21 1997

Fingerprint

Sperm Motility
Cyclic AMP-Dependent Protein Kinases
Protein Kinases
Peptides
Spermatozoa
Catalyst activity
Calcium
Isoleucine
Contraceptive Agents
Proline
Protein Binding
Cell Communication
Suspensions
Buffers
Protein Isoforms
Homeostasis
Substitution reactions
Chemical activation
Modulation
S Ht31

ASJC Scopus subject areas

  • Biochemistry

Cite this

Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility. / Vijayaraghavan, Srinivasan; Goueli, Said A.; Davey, Michael; Carr, Daniel.

In: Journal of Biological Chemistry, Vol. 272, No. 8, 21.02.1997, p. 4747-4752.

Research output: Contribution to journalArticle

Vijayaraghavan, Srinivasan ; Goueli, Said A. ; Davey, Michael ; Carr, Daniel. / Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 8. pp. 4747-4752.
@article{6a30a9f109c64896b990d08cb7900e6b,
title = "Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility",
abstract = "Cyclic AMP-dependent protein kinase (PKA) is anchored at specific subcellular sites through the interaction of the regulatory subunit (R) with protein kinase A-anchoring proteins (AKAPs) via an amphipathic helix binding motif. Synthetic peptides containing this amphipathic helix domain competitively disrupt PKA binding to AKAPs and cause a loss of PKA modulation of cellular responses. In this report we use S-Ht31, a cell-permeant anchoring inhibitor peptide, to study the role of PKA anchoring in sperm. Our analysis of three species of mammalian sperm detected three isoforms of PKA (RIIα, RIIβ, and RIβ) and one 110-kDa AKAP. The addition of S-Ht31 to bovine caudal epididymal sperm inhibits motility in a time- and concentration-dependent manner. A control peptide, S-Ht31-P, identical to S- Ht31 except for a proline for isoleucine substitution to prevent amphipathic helix formation, had no effect on motility. The inhibition of motility by S- Ht31 is reversible but only if calcium is present in the suspension buffer, suggesting a role for PKA anchoring in regulating cellular calcium homeostasis. Surprisingly, inhibition of PKA catalytic activity had little effect on basal motility or motility stimulated by agents previously thought to work via PKA activation. These data suggest that the interaction of the regulatory subunit of PKA with sperm AKAPs, independent of PKA catalytic activity, is a key regulator of sperm motility and that disruption of this interaction using cell-permeable anchoring inhibitor peptides may form the basis of a sperm-targeted contraceptive.",
author = "Srinivasan Vijayaraghavan and Goueli, {Said A.} and Michael Davey and Daniel Carr",
year = "1997",
month = "2",
day = "21",
doi = "10.1074/jbc.272.8.4747",
language = "English (US)",
volume = "272",
pages = "4747--4752",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "8",

}

TY - JOUR

T1 - Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility

AU - Vijayaraghavan, Srinivasan

AU - Goueli, Said A.

AU - Davey, Michael

AU - Carr, Daniel

PY - 1997/2/21

Y1 - 1997/2/21

N2 - Cyclic AMP-dependent protein kinase (PKA) is anchored at specific subcellular sites through the interaction of the regulatory subunit (R) with protein kinase A-anchoring proteins (AKAPs) via an amphipathic helix binding motif. Synthetic peptides containing this amphipathic helix domain competitively disrupt PKA binding to AKAPs and cause a loss of PKA modulation of cellular responses. In this report we use S-Ht31, a cell-permeant anchoring inhibitor peptide, to study the role of PKA anchoring in sperm. Our analysis of three species of mammalian sperm detected three isoforms of PKA (RIIα, RIIβ, and RIβ) and one 110-kDa AKAP. The addition of S-Ht31 to bovine caudal epididymal sperm inhibits motility in a time- and concentration-dependent manner. A control peptide, S-Ht31-P, identical to S- Ht31 except for a proline for isoleucine substitution to prevent amphipathic helix formation, had no effect on motility. The inhibition of motility by S- Ht31 is reversible but only if calcium is present in the suspension buffer, suggesting a role for PKA anchoring in regulating cellular calcium homeostasis. Surprisingly, inhibition of PKA catalytic activity had little effect on basal motility or motility stimulated by agents previously thought to work via PKA activation. These data suggest that the interaction of the regulatory subunit of PKA with sperm AKAPs, independent of PKA catalytic activity, is a key regulator of sperm motility and that disruption of this interaction using cell-permeable anchoring inhibitor peptides may form the basis of a sperm-targeted contraceptive.

AB - Cyclic AMP-dependent protein kinase (PKA) is anchored at specific subcellular sites through the interaction of the regulatory subunit (R) with protein kinase A-anchoring proteins (AKAPs) via an amphipathic helix binding motif. Synthetic peptides containing this amphipathic helix domain competitively disrupt PKA binding to AKAPs and cause a loss of PKA modulation of cellular responses. In this report we use S-Ht31, a cell-permeant anchoring inhibitor peptide, to study the role of PKA anchoring in sperm. Our analysis of three species of mammalian sperm detected three isoforms of PKA (RIIα, RIIβ, and RIβ) and one 110-kDa AKAP. The addition of S-Ht31 to bovine caudal epididymal sperm inhibits motility in a time- and concentration-dependent manner. A control peptide, S-Ht31-P, identical to S- Ht31 except for a proline for isoleucine substitution to prevent amphipathic helix formation, had no effect on motility. The inhibition of motility by S- Ht31 is reversible but only if calcium is present in the suspension buffer, suggesting a role for PKA anchoring in regulating cellular calcium homeostasis. Surprisingly, inhibition of PKA catalytic activity had little effect on basal motility or motility stimulated by agents previously thought to work via PKA activation. These data suggest that the interaction of the regulatory subunit of PKA with sperm AKAPs, independent of PKA catalytic activity, is a key regulator of sperm motility and that disruption of this interaction using cell-permeable anchoring inhibitor peptides may form the basis of a sperm-targeted contraceptive.

UR - http://www.scopus.com/inward/record.url?scp=0031040893&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031040893&partnerID=8YFLogxK

U2 - 10.1074/jbc.272.8.4747

DO - 10.1074/jbc.272.8.4747

M3 - Article

C2 - 9030527

AN - SCOPUS:0031040893

VL - 272

SP - 4747

EP - 4752

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 8

ER -