题名：Peptidomic Profiling of Secreted Products from Pancreatic Islet Culture Results in a Higher Yield of Full-length Peptide Hormones than Found using Cell Lysis Procedures.
作者：TAYLOR SW; NIKOULINA SE; ANDON NL; LOWE C;
来源：J Proteome Res. 2013 Jun 28. [ IF= 0.00 ] ]
摘要：Peptide Hormone Acquisition through Smart Sampling Technique-Mass Spectrometry
(PHASST-MS) is a peptidomics platform that employs high resolution liquid
chromatography-mass spectrometry (LC-MS) techniques to identify peptide hormones
secreted from in vitro or ex vivo cultures enriched in endocrine cells.
Application of the methodology to the study of murine pancreatic islets has
permitted evaluation of the strengths and weaknesses of the approach, as well as
comparison of our results with published islet studies that employed traditional
cellular lysis procedures. We found that, while our PHASST-MS approach identified
fewer peptides in total, we had greater representation of intact peptide
hormones. The technique was further refined to improve coverage of hydrophilic as
well as hydrophobic peptides and subsequently applied to human pancreatic islet
cultures derived from normal donors or donors with type 2 diabetes.
Interestingly, in addition to the expected islet hormones, we identified
alpha-cell-derived bioactive GLP-1, consistent with recent reports of paracrine
effects of this hormone on beta-cell function. We also identified many novel
peptides derived from neurohormonal precursors and proteins related to the cell
secretory system. Taken together, these results suggest the PHASST-MS strategy of
focusing on cellular secreted products rather than the total tissue peptidome may
improve the probability of discovering novel bioactive peptides and also has the
potential to offer important new insights into the secretion and function of