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Animal Models

Endoplasmic reticulum-associated degradation of the mouse PC1/3-N222D hypomorph and human PCSK1 mutations contributes to obesity

International Journal of Obesity volume 40, pages 973–981 (2016)Cite this article

Subjects

Abstract

Background:

The proprotein convertase 1/3 (PC1/3), encoded by proprotein convertase subtilisin/kexin type 1 ( PCSK1), cleaves and hence activates several orexigenic and anorexigenic proproteins. Congenital inactivation of PCSK1 leads to obesity in human but not in mice. However, a mouse model harboring the hypomorphic mutation N222D is obese. It is not clear why the mouse models differ in phenotype.

Methods:

Gene expression analysis was performed with pancreatic islets from Pcsk1N222D/N222D mice. Subsequently, biosynthesis, maturation, degradation and activity were studied in islets, pituitary, hypothalamus and cell lines. Coimmunoprecipitation of PC1/3-N222D and human PC1/3 variants associated with obesity with the endoplasmic reticulum (ER) chaperone BiP was studied in cell lines.

Results:

Gene expression analysis of islets of Pcsk1N222D/N222D mice showed enrichment of gene sets related to the proteasome and the unfolded protein response. Steady-state levels of PC1/3-N222D and in particular the carboxy-terminally processed form were strongly reduced in islets, pituitary and hypothalamus. However, impairment of substrate cleavage was tissue dependent. Proinsulin processing was drastically reduced, while processing of proopiomelanocortin (POMC) to adrenocorticotropic hormone (ACTH) in pituitary was only mildly impaired. Growth hormone expression and IGF-1 levels were normal, indicating near-normal processing of hypothalamic proGHRH. PC1/3-N222D binds to BiP and is rapidly degraded by the proteasome. Analysis of human PC1/3 obesity-associated mutations showed increased binding to BiP and prolonged intracellular retention for all investigated mutations, in particular for PC1/3-T175M, PC1/3-G226R and PC1/3-G593R.

Conclusions:

This study demonstrates that the hypomorphic mutation in Pcsk1N222D mice has an effect on catalytic activity in pancreatic islets, pituitary and hypothalamus. Reduced substrate processing activity in Pcsk1N222D/N222D mice is due to enhanced degradation in addition to reduced catalytic activity of the mutant. PC1/3-N222D binds to BiP, suggesting impaired folding and reduced stability. Enhanced BiP binding is also observed in several human obesity-associated PC1/3 variants, suggesting a common mechanism.

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Acknowledgements

PS designed the study, researched data and wrote the manuscript. BB researched data and reviewed the manuscript. ED and LV researched data, BR, FS, LT and JD reviewed the manuscript. JWMC wrote the manuscript and is guarantor of this article. This work was supported by IWT-Vlaanderen, personal fellowship PS, FWO Vlaanderen and KU Leuven grant GOA2008/16. We wish to thank Sandra Meulemans for technical assistance, Fred Van Leuven for advice and Robert Day, Ann White and Iris Lindberg for providing materials.

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Authors and Affiliations

  1. Department of Human Genetics, Laboratory for Biochemical Neuroendocrinology, KU Leuven, Leuven,, Belgium

    P Stijnen, B Brouwers, E Dirkx, B Ramos-Molina, J Declercq & J W M Creemers

  2. Department of Cellular and Molecular Medicine, Gene Expression Unit, KU Leuven, Leuven,, Belgium

    L Van Lommel & F Schuit

  3. Department of Development and Regeneration, KU Leuven, Leuven,, Belgium

    L Thorrez

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Stijnen, P., Brouwers, B., Dirkx, E. et al. Endoplasmic reticulum-associated degradation of the mouse PC1/3-N222D hypomorph and human PCSK1 mutations contributes to obesity. Int J Obes 40, 973–981 (2016). https://doi.org/10.1038/ijo.2016.3

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