Cushing's syndrome mutant PKAL205R exhibits altered substrate specificity

Lubner, Joshua M.; Dodge-Kafka, Kimberly L.; Carlson, Cathrine R.; Church, George M.; Chou, Michael F.; Schwartz, Daniel

doi:10.1002/1873-3468.12562

Title: Cushing's syndrome mutant PKA^L205R exhibits altered substrate specificity

Abstract

The PKA ^L ^205R hotspot mutation has been implicated in Cushing's syndrome through hyperactive gain‐of‐function PKA signaling; however, its influence on substrate specificity has not been investigated. Here, we employ the Proteomic Peptide Library (ProPeL) approach to create high‐resolution models for PKA ^WT and PKA ^L ^205R substrate specificity. We reveal that the L205R mutation reduces canonical hydrophobic preference at the substrate P + 1 position, and increases acidic preference in downstream positions. Using these models, we designed peptide substrates that exhibit altered selectivity for specific PKA variants, and demonstrate the feasibility of selective PKA ^L ^205R loss‐of‐function signaling. Through these results, we suggest that substrate rewiring may contribute to Cushing's syndrome disease etiology, and introduce a powerful new paradigm for investigating mutation‐induced kinase substrate rewiring in human disease.

Authors:

Lubner, Joshua M. ^[1]; Dodge-Kafka, Kimberly L. ^[2]; Carlson, Cathrine R. ^[3]; Church, George M. ^[4]; Chou, Michael F. ^[4]; Schwartz, Daniel ^[1]

Univ. of Connecticut, Storrs, CT (United States). Dept. of Physiology and Neurobiology
Univ. of Connecticut Health Center, Farmington CT (United States). Pat and Jim Calhoun Center for Cardiology, Dept. of Cell Biology
Oslo University Hospital and University of Oslo (Norway). Inst. for Experimental Medical Research
Harvard Medical School, Boston, MA (United States). Dept. of Genetics; Harvard Univ., Boston, MA (United States). Wyss Inst. For Biologically Inspired Engineering

Publication Date:: Wed Feb 01 00:00:00 EST 2017

Research Org.:: Harvard Univ., Cambridge, MA (United States)

Sponsoring Org.:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH), National Institute Of Neurological Disorders and Stroke; University of Connecticut Research Foundation

OSTI Identifier:: 1345186

Alternate Identifier(s):: OSTI ID: 1399283

Grant/Contract Number:: FG02-02ER63445; DE‐FG02‐02ER63445

Resource Type:: Accepted Manuscript

Journal Name:: FEBS Letters

Additional Journal Information:: Journal Volume: 591; Journal Issue: 3; Related Information: Supplementary information files are available on the journal web site: http://onlinelibrary.wiley.com/doi/10.1002/1873-3468.12562/full in the Supporting Information section. They comprise three tables of proteomic and in vitro-derived data and an Appendix to the article.; Journal ID: ISSN 0014-5793

Publisher:: Federation of European Biochemical Societies

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; Cushing's syndrome; protein kinase A; substrate specificity; proteomic peptide library

Citation Formats


                    Lubner, Joshua M., Dodge-Kafka, Kimberly L., Carlson, Cathrine R., Church, George M., Chou, Michael F., and Schwartz, Daniel. Cushing's syndrome mutant PKAL205R exhibits altered substrate specificity.  United States: N. p., 2017. 
Web.  doi:10.1002/1873-3468.12562.

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                    Lubner, Joshua M., Dodge-Kafka, Kimberly L., Carlson, Cathrine R., Church, George M., Chou, Michael F., & Schwartz, Daniel. Cushing's syndrome mutant PKAL205R exhibits altered substrate specificity.  United States.  https://doi.org/10.1002/1873-3468.12562

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                    Lubner, Joshua M., Dodge-Kafka, Kimberly L., Carlson, Cathrine R., Church, George M., Chou, Michael F., and Schwartz, Daniel. Wed .  
"Cushing's syndrome mutant PKAL205R exhibits altered substrate specificity".  United States.  https://doi.org/10.1002/1873-3468.12562.  https://www.osti.gov/servlets/purl/1345186.

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@article{osti_1345186,

  title        = {Cushing's syndrome mutant PKAL205R exhibits altered substrate specificity},

  author       = {Lubner, Joshua M. and Dodge-Kafka, Kimberly L. and Carlson, Cathrine R. and Church, George M. and Chou, Michael F. and Schwartz, Daniel},

  abstractNote = {The PKA L 205R hotspot mutation has been implicated in Cushing's syndrome through hyperactive gain‐of‐function PKA signaling; however, its influence on substrate specificity has not been investigated. Here, we employ the Proteomic Peptide Library (ProPeL) approach to create high‐resolution models for PKA WT and PKA L 205R substrate specificity. We reveal that the L205R mutation reduces canonical hydrophobic preference at the substrate P + 1 position, and increases acidic preference in downstream positions. Using these models, we designed peptide substrates that exhibit altered selectivity for specific PKA variants, and demonstrate the feasibility of selective PKA L 205R loss‐of‐function signaling. Through these results, we suggest that substrate rewiring may contribute to Cushing's syndrome disease etiology, and introduce a powerful new paradigm for investigating mutation‐induced kinase substrate rewiring in human disease.},

  doi          = {10.1002/1873-3468.12562},

  journal      = {FEBS Letters},

  number       = 3,

  volume       = 591,

  place        = {United States},

  year         = {Wed Feb 01 00:00:00 EST 2017},

  month        = {Wed Feb 01 00:00:00 EST 2017}

}

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https://doi.org/10.1002/1873-3468.12562

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