Top-Down Proteomics of Mouse Islets With Beta Cell CPE Deletion Reveals Molecular Details in Prohormone Processing

Fulcher, James M.; Swensen, Adam C.; Chen, Yi-Chun; Verchere, C. Bruce; Petyuk, Vladislav A.; Qian, Weijun

doi:10.1210/endocr/bqad160

Top-Down Proteomics of Mouse Islets With Beta Cell CPE Deletion Reveals Molecular Details in Prohormone Processing

Journal Article · Wed Nov 15 04:00:00 EST 2023 · Endocrinology (Online)

DOI:https://doi.org/10.1210/endocr/bqad160· OSTI ID:2331436

Fulcher, James M. ^[1]; Swensen, Adam C. ^[2]; Chen, Yi-Chun ^[3]; Verchere, C. Bruce ^[3]; ^[2]; ^[2]

Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)
Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
University of British Columbia, Vancouver, BC (Canada); BC Children’s Hospital Research Institute, Vancouver, BC (Canada)

Altered prohormone processing, such as with proinsulin and pro-islet amyloid polypeptide (proIAPP), has been reported as an important feature of prediabetes and diabetes. Proinsulin processing includes removal of several C-terminal basic amino acids and is performed principally by the exopeptidase carboxypeptidase E (CPE), and mutations in CPE or other prohormone convertase enzymes (PC1/3 and PC2) result in hyperproinsulinemia. A comprehensive characterization of the forms and quantities of improperly processed insulin and other hormone products following Cpe deletion in pancreatic islets has yet to be attempted. In the present study we applied top-down proteomics to globally evaluate the numerous proteoforms of hormone processing intermediates in a β-cell-specific Cpe knockout mouse model. Increases in dibasic residue–containing proinsulin and other novel proteoforms of improperly processed proinsulin were found, and we could classify several processed proteoforms as novel substrates of CPE. Interestingly, some other known substrates of CPE remained unaffected despite its deletion, implying that paralogous processing enzymes such as carboxypeptidase D (CPD) can compensate for CPE loss and maintain near normal levels of hormone processing. In summary, our quantitative results from top-down proteomics of islets provide unique insights into the complexity of hormone processing products and the regulatory mechanisms.

View Accepted Manuscript (DOE)

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); Juvenile Diabetes Research Federation; Canadian Institutes of Health and Research

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 2331436

Report Number(s):: PNNL-SA--188096

Journal Information:: Endocrinology (Online), Journal Name: Endocrinology (Online) Journal Issue: 12 Vol. 164; ISSN 1945-7170

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Top-Down Proteomics of Mouse Islets With Beta Cell CPE Deletion Reveals Molecular Details in Prohormone Processing

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