Bioinformatic design of A-kinase anchoring protein-in silico: A potent and selective peptide antagonist of type II protein kinase A anchoring

Neal M. Alto, Scott H. Soderling, Naoto Hoshi, Lorene K. Langeberg, Rosa Fayos, Patricia A. Jennings, John D. Scott

Research output: Contribution to journalArticle

136 Citations (Scopus)

Abstract

Compartmentalization of the cAMP-dependent protein kinase (PKA) is coordinated through association with A-kinase anchoring proteins (AKAPs). A defining characteristic of most AKAPs is a 14-to 18-aa sequence that binds to the regulatory subunits (RI or RII) of the kinase. Cellular delivery of peptides to these regions disrupts PKA anchoring and has been used to delineate a physiological role for AKAPs in the facilitation of certain cAMP-responsive events. Here, we describe a bioinformatic approach that yields an RII-selective peptide, called AKAP-in silico (AKAP-IS), that binds RII with a Kd of 0.4 nM and binds RI with a Kd of 277 nM. AKAP-IS associates with the type II PKA holoenzyme inside cells and displaces the kinase from natural anchoring sites. Electrophysiological recordings indicate that perfusion of AKAP-IS evokes a more rapid and complete attenuation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents than previously described anchoring inhibitor peptides. Thus, computer-based and peptide array screening approaches have generated a reagent that binds PKA with higher affinity than previously described AKAPs.

Original languageEnglish (US)
Pages (from-to)4445-4450
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume100
Issue number8
DOIs
StatePublished - Apr 15 2003

Fingerprint

Cyclic AMP-Dependent Protein Kinase Type II
Computational Biology
Computer Simulation
Protein Kinases
Peptides
Phosphotransferases
Holoenzymes
AMPA Receptors
Cyclic AMP-Dependent Protein Kinases

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Bioinformatic design of A-kinase anchoring protein-in silico : A potent and selective peptide antagonist of type II protein kinase A anchoring. / Alto, Neal M.; Soderling, Scott H.; Hoshi, Naoto; Langeberg, Lorene K.; Fayos, Rosa; Jennings, Patricia A.; Scott, John D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 8, 15.04.2003, p. 4445-4450.

Research output: Contribution to journalArticle

Alto, Neal M. ; Soderling, Scott H. ; Hoshi, Naoto ; Langeberg, Lorene K. ; Fayos, Rosa ; Jennings, Patricia A. ; Scott, John D. / Bioinformatic design of A-kinase anchoring protein-in silico : A potent and selective peptide antagonist of type II protein kinase A anchoring. In: Proceedings of the National Academy of Sciences of the United States of America. 2003 ; Vol. 100, No. 8. pp. 4445-4450.
@article{156c2d84a81c47ef8be8574f7ae970f9,
title = "Bioinformatic design of A-kinase anchoring protein-in silico: A potent and selective peptide antagonist of type II protein kinase A anchoring",
abstract = "Compartmentalization of the cAMP-dependent protein kinase (PKA) is coordinated through association with A-kinase anchoring proteins (AKAPs). A defining characteristic of most AKAPs is a 14-to 18-aa sequence that binds to the regulatory subunits (RI or RII) of the kinase. Cellular delivery of peptides to these regions disrupts PKA anchoring and has been used to delineate a physiological role for AKAPs in the facilitation of certain cAMP-responsive events. Here, we describe a bioinformatic approach that yields an RII-selective peptide, called AKAP-in silico (AKAP-IS), that binds RII with a Kd of 0.4 nM and binds RI with a Kd of 277 nM. AKAP-IS associates with the type II PKA holoenzyme inside cells and displaces the kinase from natural anchoring sites. Electrophysiological recordings indicate that perfusion of AKAP-IS evokes a more rapid and complete attenuation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents than previously described anchoring inhibitor peptides. Thus, computer-based and peptide array screening approaches have generated a reagent that binds PKA with higher affinity than previously described AKAPs.",
author = "Alto, {Neal M.} and Soderling, {Scott H.} and Naoto Hoshi and Langeberg, {Lorene K.} and Rosa Fayos and Jennings, {Patricia A.} and Scott, {John D.}",
year = "2003",
month = "4",
day = "15",
doi = "10.1073/pnas.0330734100",
language = "English (US)",
volume = "100",
pages = "4445--4450",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "8",

}

TY - JOUR

T1 - Bioinformatic design of A-kinase anchoring protein-in silico

T2 - A potent and selective peptide antagonist of type II protein kinase A anchoring

AU - Alto, Neal M.

AU - Soderling, Scott H.

AU - Hoshi, Naoto

AU - Langeberg, Lorene K.

AU - Fayos, Rosa

AU - Jennings, Patricia A.

AU - Scott, John D.

PY - 2003/4/15

Y1 - 2003/4/15

N2 - Compartmentalization of the cAMP-dependent protein kinase (PKA) is coordinated through association with A-kinase anchoring proteins (AKAPs). A defining characteristic of most AKAPs is a 14-to 18-aa sequence that binds to the regulatory subunits (RI or RII) of the kinase. Cellular delivery of peptides to these regions disrupts PKA anchoring and has been used to delineate a physiological role for AKAPs in the facilitation of certain cAMP-responsive events. Here, we describe a bioinformatic approach that yields an RII-selective peptide, called AKAP-in silico (AKAP-IS), that binds RII with a Kd of 0.4 nM and binds RI with a Kd of 277 nM. AKAP-IS associates with the type II PKA holoenzyme inside cells and displaces the kinase from natural anchoring sites. Electrophysiological recordings indicate that perfusion of AKAP-IS evokes a more rapid and complete attenuation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents than previously described anchoring inhibitor peptides. Thus, computer-based and peptide array screening approaches have generated a reagent that binds PKA with higher affinity than previously described AKAPs.

AB - Compartmentalization of the cAMP-dependent protein kinase (PKA) is coordinated through association with A-kinase anchoring proteins (AKAPs). A defining characteristic of most AKAPs is a 14-to 18-aa sequence that binds to the regulatory subunits (RI or RII) of the kinase. Cellular delivery of peptides to these regions disrupts PKA anchoring and has been used to delineate a physiological role for AKAPs in the facilitation of certain cAMP-responsive events. Here, we describe a bioinformatic approach that yields an RII-selective peptide, called AKAP-in silico (AKAP-IS), that binds RII with a Kd of 0.4 nM and binds RI with a Kd of 277 nM. AKAP-IS associates with the type II PKA holoenzyme inside cells and displaces the kinase from natural anchoring sites. Electrophysiological recordings indicate that perfusion of AKAP-IS evokes a more rapid and complete attenuation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents than previously described anchoring inhibitor peptides. Thus, computer-based and peptide array screening approaches have generated a reagent that binds PKA with higher affinity than previously described AKAPs.

UR - http://www.scopus.com/inward/record.url?scp=0037447227&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037447227&partnerID=8YFLogxK

U2 - 10.1073/pnas.0330734100

DO - 10.1073/pnas.0330734100

M3 - Article

C2 - 12672969

AN - SCOPUS:0037447227

VL - 100

SP - 4445

EP - 4450

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 8

ER -