title: Sample preparation protocol for bottom-up proteomic analysis of the secretome of the islets of Langerhans.
abstract: We evaluate a set of protocols for preparation of the secretome from murine islets of Langerhans for bottom-up proteomic analysis. Of the protocols evaluated, a filter-aided sample preparation based approach using sodium dodecyl sulfate as a detergent to solubilize proteins generated the most protein identifications. A total of 362 protein groups (average of 3.7 peptides/protein) were identified from the secretome using the SDS-FASP protocol; a combination of data from three protocols generated 413 protein group identifications. As expected, the identified proteins included insulin 1 and 2, somatostatin, and glucagon, the four main secreted components from islets. STRING network analysis classified the other proteins as being associated with extracellular exosomes, membrane-bounded vesicles, vesicles, and the extracellular region.
authors: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA. ndovichi@nd.edu.
authors: Andrew
authors: A
authors: Schmudlach
authors: Departments of Chemistry and Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA.
authors: Jeremy
authors: J
authors: Felton
authors: Departments of Chemistry and Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA.
authors: Cynthia
authors: C
authors: Cipolla
authors: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA. ndovichi@nd.edu.
authors: Liangliang
authors: L
authors: Sun
authors: Departments of Chemistry and Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA.
authors: Robert T
authors: RT
authors: Kennedy
authors: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA. ndovichi@nd.edu.
authors: Norman J
authors: NJ
authors: Dovichi
mesh_terms: D053000:Analytic Sample Preparation Methods / Q000379:methods*; D000818:Animals; D007515:Islets of Langerhans / Q000378:metabolism*; D051379:Mice; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics
chemical_list: D011506:Proteins