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Title: Cushing's syndrome mutant PKAL205R exhibits altered substrate specificity

Journal Article · · FEBS Letters
 [1];  [2];  [3];  [4];  [4];  [1]
  1. Univ. of Connecticut, Storrs, CT (United States). Dept. of Physiology and Neurobiology
  2. Univ. of Connecticut Health Center, Farmington CT (United States). Pat and Jim Calhoun Center for Cardiology, Dept. of Cell Biology
  3. Oslo University Hospital and University of Oslo (Norway). Inst. for Experimental Medical Research
  4. Harvard Medical School, Boston, MA (United States). Dept. of Genetics; Harvard Univ., Boston, MA (United States). Wyss Inst. For Biologically Inspired Engineering
+ Show Author Affiliations

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.

Research Organization:
Harvard Univ., Cambridge, MA (United States)
Sponsoring Organization:
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
Grant/Contract Number:
FG02-02ER63445; DE‐FG02‐02ER63445
OSTI ID:
1345186
Alternate ID(s):
OSTI ID: 1399283
Journal Information:
FEBS Letters, Vol. 591, 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.; ISSN 0014-5793
Publisher:
Federation of European Biochemical SocietiesCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 14 works
Citation information provided by
Web of Science

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Cited By (5)

Kinetics and inhibition studies of the L205R mutant of cAMP ‐dependent protein kinase involved in Cushing's syndrome journal March 2018
Cushing’s syndrome driver mutation disrupts protein kinase A allosteric network, altering both regulation and substrate specificity journal August 2019
Alterations in Protein Kinase A Substrate Specificity as a Potential Cause of Cushing Syndrome journal January 2019
Evolution of protein kinase substrate recognition at the active site journal June 2019
Evolution of protein kinase substrate recognition at the active site posted_content October 2018

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