Mutations in the PCSK1 gene encoding
prohormone convertase 1/3 (PC1/3) are strongly associated with
obesity in humans. The PC1/3(N222D) mutant mouse thus far represents the only mouse model that mimics the PC1/3
obesity phenotype in humans. The present investigation addresses the cell biology of the N222D mutation. Metabolic labeling experiments reveal a clear defect in the kinetics of
insulin biosynthesis in islets from PC1/3(N222D) mutant mice, resulting in an increase in both
proinsulin and its processing intermediates, predominantly lacking cleavage at the
Arg-Arg site. Although the mutant PC1/3
zymogen is correctly processed to the 87-kDa form, pulse-chase immunoprecipitation experiments, labeling, and immunohistochemical experiments using uncleavable variants all demonstrate that the PC1/3-N222D
protein is largely mislocalized compared with similar wild-type (WT) constructs, being predominantly retained in the endoplasmic reticulum. The PC1/3-N222D mutant also undergoes more efficient degradation via the
ubiquitin-
proteasome system than the WT
enzyme. Lastly, the mutant PC1/3-N222D
protein coimmunoprecipitates with WT PC1/3 and exerts a modest effect on intracellular retention of the WT
enzyme. These profound alterations in the cell biology of PC1/3-N222D are likely to contribute to the defective
insulin biosynthetic events observed in the mutant mice and may be relevant to the dramatic contributions of polymorphisms in this gene to human
obesity.