Apoe, Mbl2, and Psp Plasma Protein Levels Correlate with Diabetic Phenotype in NZO MiceAn Optimized Rapid Workflow for SRM-Based Quantification

• Published in: Journal of proteome research 2013-03, Vol.12 (3), p.1331-1343
• Main Authors: von Toerne, Christine, Kahle, Melanie, Schäfer, Alexander, Ispiryan, Ruben, Blindert, Marcel, Hrabe De Angelis, Martin, Neschen, Susanne, Ueffing, Marius, Hauck, Stefanie M
• Format: Article
• Language: English
• Subjects: Animals; Apolipoproteins E - blood; Chromatography, High Pressure Liquid; Diabetes Mellitus, Experimental - blood; Enzyme-Linked Immunosorbent Assay; Male; Mannose-Binding Lectin - blood; Mice; Mice, Obese; Phenotype; Proteomics; Reproducibility of Results; Salivary Proteins and Peptides - blood; Tandem Mass Spectrometry
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr3009836

Male New Zealand Obese (NZO) mice progress through pathophysiological stages similar to humans developing obesity-associated type 2 diabetes (T2D). The current challenge is to establish quantitative proteomics from small plasma sample amounts. We established an analytical workflow that facilitates a reproducible depletion of high-abundance proteins, has high throughput applicability, and allows absolute quantification of proteins from mouse plasma samples by LC–SRM-MS. The ProteoMiner equalizing technology was adjusted to the small sample amount, and reproducibility of the identifications was monitored by spike proteins. Based on the label-free relative quantification of proteins in depleted plasma of a test set of NZO mice, assays for potential candidates were designed for the setup of a targeted selected reaction monitoring (SRM) approach and absolute quantification. We could demonstrate that apolipoprotein E (Apoe), mannose-binding lectin 2 (Mbl2), and parotid secretory protein (Psp) are present at significantly different quantities in depleted plasma of diabetic NZO mice compared to non-diabetic controls using AQUA peptides. Quantification was validated for Mbl2 using the ELISA technology on non-depleted plasma. We conclude that the depletion technique is applicable to restricted sample amounts and suitable for the identification of T2D signatures in plasma.