Summary
Pancreatic zymogen granules (ZGs) are specialized for digestive enzyme storage and regulated secretion in exocrine pancreas and are a classical model for studying secretory granule function. To understand the function of this organelle, we have conducted a proteomic study to identify the ZG membrane (ZGM) proteins from ZGs purified by Percoll gradient centrifugation. By combining multiple separation strategies including two-dimensional gel electrophoresis and two-dimensional HPLC with tandem mass spectrometry, we identified 101 proteins from purified ZGMs including a large number of proteins previously unknown on ZGMs. To distinguish intrinsic membrane proteins from soluble and peripheral membrane proteins, a quantitative proteomics strategy was used to measure the enrichment of intrinsic membrane proteins through the purification steps by labeling crude, KBr-, and Na\(_{2}\)CO\(_{3}\)-washed ZGMs with multiplexed isobaric tags (iTRAQ™), 114, 116, and 117, respectively. The proteins with 117:114 ratios greater than one correlated well with intrinsic membrane proteins that contain either known or predicted transmembrane domains.
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Acknowledgments
We gratefully acknowledge Dr. John A. Williams for helping us to prepare the manuscript. We thank the Michigan Proteome Consortium for performing some of the analyses. This research was supported by the National Resource for Proteomics and Pathways (P41 RR018627) to Philip C. Andrews and also by NIH grants DK 41122 to John A. Williams.
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Chen, X., Andrews, P.C. (2008). Purification and Proteomics Analysis of Pancreatic Zymogen Granule Membranes. In: Pflieger, D., Rossier, J. (eds) Organelle Proteomics. Methods in Molecular Biology™, vol 432. Humana Press. https://doi.org/10.1007/978-1-59745-028-7_19
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DOI: https://doi.org/10.1007/978-1-59745-028-7_19
Publisher Name: Humana Press
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