Molecular architecture of mouse and human pancreatic zymogen granules: protein components and their copy numbers

Jin-sook Lee; Joseph A. Caruso; Garrett Hubbs; Patricia Schnepp; James Woods; Jingye Fang; Chunying Li; Kezhong Zhang; Paul M. Stemmer; Bhanu P. Jena; Xuequn Chen

doi:10.1007/s41048-018-0055-1

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Article Navigation > Biophysics Reports > 2018 > 4(2): 94-103

Jin-sook Lee, Joseph A. Caruso, Garrett Hubbs, Patricia Schnepp, James Woods, Jingye Fang, Chunying Li, Kezhong Zhang, Paul M. Stemmer, Bhanu P. Jena, Xuequn Chen. Molecular architecture of mouse and human pancreatic zymogen granules: protein components and their copy numbers. Biophysics Reports, 2018, 4(2): 94-103. doi: 10.1007/s41048-018-0055-1

Citation:

Jin-sook Lee, Joseph A. Caruso, Garrett Hubbs, Patricia Schnepp, James Woods, Jingye Fang, Chunying Li, Kezhong Zhang, Paul M. Stemmer, Bhanu P. Jena, Xuequn Chen. Molecular architecture of mouse and human pancreatic zymogen granules: protein components and their copy numbers. Biophysics Reports, 2018, 4(2): 94-103. doi: 10.1007/s41048-018-0055-1

Citation:

Jin-sook Lee, Joseph A. Caruso, Garrett Hubbs, Patricia Schnepp, James Woods, Jingye Fang, Chunying Li, Kezhong Zhang, Paul M. Stemmer, Bhanu P. Jena, Xuequn Chen. Molecular architecture of mouse and human pancreatic zymogen granules: protein components and their copy numbers. Biophysics Reports, 2018, 4(2): 94-103. doi: 10.1007/s41048-018-0055-1

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Molecular architecture of mouse and human pancreatic zymogen granules: protein components and their copy numbers

doi: 10.1007/s41048-018-0055-1

1 Department of Physiology, Wayne State University, Detroit, MI 48201, USA
2 School of Medicine, Institute of Environmental Health Sciences, Wayne State University, Detroit, MI 48201, USA
3 Department of Biochemistry and Molecular Biology, Wayne State University, Detroit, MI 48201, USA
4 Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI 48201, USA

Funds: This work is supported by the start-up fund from Wayne State University and partly by National Institute of Diabetes and Digestive and Kidney Diseases Grant (1R01DK110314).We sincerely thank Dr.Zong Jie Cui for reading our manuscript and his insightful comments.We also acknowledge the assistance of the Wayne State University Proteomics Core that is supported through NIH grants P30 ES020957,P30 CA 022453,and S10 OD010700.

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Corresponding author: Xuequn Chen
Received Date: 25 April 2017
Rev Recd Date: 17 October 2017
Publish Date: 30 April 2018

Abstract

Abstract

A molecular model of pancreatic zymogen granule (ZG) is critical for understanding its functions. We have extensively characterized the composition and membrane topology of rat ZG proteins. In this study, we report the development of targeted proteomics approaches to quantify representative mouse and human ZG proteins using LC-SRM and heavy isotope-labeled synthetic peptides. The absolute quantities of mouse Rab3D and VAMP8 were determined as 1242 ±218 and 2039 ±151 (mean ±SEM) copies per ZG. The size distribution and the averaged diameter of ZGs 750 ±23 nm (mean ±SEM) were determined by atomic force microscopy. The absolute quantification of Rab3D was then validated using semi-quantitative Western blotting with purified GST-Rab3D proteins as an internal standard. To extend our proteomics analysis to human pancreas, ZGs were purified using human acini obtained from pancreatic islet transplantation center. One hundred and eighty human ZG proteins were identified for the first time including both the membrane and the content proteins. Furthermore, the copy number per ZG of human Rab3D and VAMP8 were determined to be 1182 ±45 and 485 ±15 (mean ±SEM). The comprehensive proteomic analyses of mouse and human pancreatic ZGs have the potential to identify species-specific ZG proteins. The determination of protein copy numbers on pancreatic ZGs represents a significant advance towards building a quantitative molecular model of a prototypical secretory vesicle using targeted proteomics approaches. The identification of human ZG proteins lays a foundation for subsequent studies of altered ZG compositions and secretion in pancreatic diseases.
- Pancreatic zymogen granule,
- GeLC-MS/MS,
- LC-SRM,
- Absolute quantification,
- Copy numbers

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References(33)

References

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