Local protein kinase A action proceeds through intact holoenzymes

F. Donelson  Smith; J.L. Esseltine; P.J. Nygren; D. Veesler; D.P. Byrne; M. Vonderach; I. Strashnov; C.E. Eyers; P.A. Eyers; L.K. Langeberg; J.D. Scott

doi:10.1126/science.aaj1669

Local protein kinase A action proceeds through intact holoenzymes

F. Donelson Smith
, J.L. Esseltine
, P.J. Nygren
, D. Veesler
, D.P. Byrne
, M. Vonderach
, I. Strashnov
, C.E. Eyers
, P.A. Eyers
, L.K. Langeberg
, J.D. Scott

Research output: Contribution to journal › Article › peer-review

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Abstract

Hormones can transmit signals through adenosine 3ʹ,5ʹ-monophosphate (cAMP) to precise intracellular locations. The fidelity of these responses relies on the activation of localized protein kinase A (PKA) holoenzymes. Association of PKA regulatory type II (RII) subunits with A-kinase–anchoring proteins (AKAPs) confers location, and catalytic (C) subunits phosphorylate substrates. Single-particle electron microscopy demonstrated that AKAP79 constrains RII-C subassemblies within 150 to 250 angstroms of its targets. Native mass spectrometry established that these macromolecular assemblies incorporated stoichiometric amounts of cAMP. Chemical-biology– and live cell–imaging techniques revealed that catalytically active PKA holoenzymes remained intact within the cytoplasm. These findings indicate that the parameters of anchored PKA holoenzyme action are much more restricted than originally anticipated.

Original language	English
Pages (from-to)	1288-1293
Number of pages	6
Journal	Science
Volume	356
Issue number	6344
DOIs	https://doi.org/10.1126/science.aaj1669
Publication status	Published - 23 Jun 2017

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10.1126/science.aaj1669

Smth et al 2-16-17 FInal combinedAccepted author manuscript, 6.93 MB

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title = "Local protein kinase A action proceeds through intact holoenzymes",

abstract = "Hormones can transmit signals through adenosine 3ʹ,5ʹ-monophosphate (cAMP) to precise intracellular locations. The fidelity of these responses relies on the activation of localized protein kinase A (PKA) holoenzymes. Association of PKA regulatory type II (RII) subunits with A-kinase–anchoring proteins (AKAPs) confers location, and catalytic (C) subunits phosphorylate substrates. Single-particle electron microscopy demonstrated that AKAP79 constrains RII-C subassemblies within 150 to 250 angstroms of its targets. Native mass spectrometry established that these macromolecular assemblies incorporated stoichiometric amounts of cAMP. Chemical-biology– and live cell–imaging techniques revealed that catalytically active PKA holoenzymes remained intact within the cytoplasm. These findings indicate that the parameters of anchored PKA holoenzyme action are much more restricted than originally anticipated.",

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year = "2017",

month = jun,

day = "23",

doi = "10.1126/science.aaj1669",

language = "English",

volume = "356",

pages = "1288--1293",

journal = "Science",

issn = "0036-8075",

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TY - JOUR

T1 - Local protein kinase A action proceeds through intact holoenzymes

AU - Smith, F. Donelson

AU - Esseltine, J.L.

AU - Nygren, P.J.

AU - Veesler, D.

AU - Byrne, D.P.

AU - Vonderach, M.

AU - Strashnov, I.

AU - Eyers, C.E.

AU - Eyers, P.A.

AU - Langeberg, L.K.

AU - Scott, J.D.

PY - 2017/6/23

Y1 - 2017/6/23

N2 - Hormones can transmit signals through adenosine 3ʹ,5ʹ-monophosphate (cAMP) to precise intracellular locations. The fidelity of these responses relies on the activation of localized protein kinase A (PKA) holoenzymes. Association of PKA regulatory type II (RII) subunits with A-kinase–anchoring proteins (AKAPs) confers location, and catalytic (C) subunits phosphorylate substrates. Single-particle electron microscopy demonstrated that AKAP79 constrains RII-C subassemblies within 150 to 250 angstroms of its targets. Native mass spectrometry established that these macromolecular assemblies incorporated stoichiometric amounts of cAMP. Chemical-biology– and live cell–imaging techniques revealed that catalytically active PKA holoenzymes remained intact within the cytoplasm. These findings indicate that the parameters of anchored PKA holoenzyme action are much more restricted than originally anticipated.

AB - Hormones can transmit signals through adenosine 3ʹ,5ʹ-monophosphate (cAMP) to precise intracellular locations. The fidelity of these responses relies on the activation of localized protein kinase A (PKA) holoenzymes. Association of PKA regulatory type II (RII) subunits with A-kinase–anchoring proteins (AKAPs) confers location, and catalytic (C) subunits phosphorylate substrates. Single-particle electron microscopy demonstrated that AKAP79 constrains RII-C subassemblies within 150 to 250 angstroms of its targets. Native mass spectrometry established that these macromolecular assemblies incorporated stoichiometric amounts of cAMP. Chemical-biology– and live cell–imaging techniques revealed that catalytically active PKA holoenzymes remained intact within the cytoplasm. These findings indicate that the parameters of anchored PKA holoenzyme action are much more restricted than originally anticipated.

UR - https://www.scopus.com/pages/publications/85021306691

U2 - 10.1126/science.aaj1669

DO - 10.1126/science.aaj1669

M3 - Article

SN - 0036-8075

VL - 356

SP - 1288

EP - 1293

JO - Science

JF - Science

IS - 6344

ER -

Local protein kinase A action proceeds through intact holoenzymes

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A Core Facility for Physical and Elemental Characterisation

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Local protein kinase A action proceeds through intact holoenzymes

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A Core Facility for Physical and Elemental Characterisation

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