An INS-1 beta-cell proteome highlights the role of fatty acid biosynthesis in glucose-stimulated insulin secretion

Abstract

Pancreatic beta cells secrete insulin as a response to rising glucose level, a process known as glucose-stimulated insulin secretion (GSIS). In this study, we used liquid chromatography tandem mass spectrometry and data-independent acquisition to acquire proteomes of rat pancreatic INS-1 832/13 beta cells that were short-term stimulated with glucose concentrations ranging from 0 to 20 mM, quantifying the behavior of 3703 proteins across 11 concentrations. Ensemble clustering of proteome profiles revealed unique response patterns of proteins expressed by INS-1 cells. 237 proteins, amongst them proteins associated with vesicular SNARE interactions, protein export, and pancreatic secretion showed an increase in abundance upon glucose stimulation, whilst the majority of proteins, including those associated with metabolic pathways such as glycolysis, the TCA cycle and the respiratory chain, did not respond to rising glucose concentrations. Interestingly, we observe that enzymes participating in fatty acid metabolism, responded distinctly, showing a "switch-on" response upon release of glucose starvation with no further changes in abundance upon increasing glucose levels. We speculate that increased activity of fatty acid metabolic activity might either be part of GSIS by replenishing membrane lipids required for vesicle-mediated exocytosis and/or by providing an electron sink to compensate for the increase in glucose catabolism. Significance of the StudyWe used high-throughput proteomics to capture comprehensive proteome changes 30 minutes post stimulation in the INS-1 832/13 beta cell line. Our study provides insights into the metabolic regulation of glucose-stimulated insulin secretion in pancreatic beta cells, specifically highlighting the early role of fatty acid biosynthesis. These findings suggest a necessary shift in focus from electrochemical to metabolic mechanisms in understanding GSIS, paving the way for future research. As the first to document proteome alterations in the initial phase of GSIS, our study furthermore documents the extent of protein abundance variability when obtaining data after short stimulation times, and therefore highlights the necessity of well-controlled study design and biological replicates. The recorded data set complements existing metabolomic and transcriptomic studies, providing a valuable resource for subsequent investigations.