Abstract
The extracellular matrix (ECM) is unique to each tissue and capable of guiding cell differentiation, migration, morphology, and function. The ECM proteome of different developmental stages has not been systematically studied in the human pancreas. In this study, we apply mass spectrometry-based quantitative proteomics strategies using N,N-dimethyl leucine isobaric tags to delineate proteome-wide and ECM-specific alterations in four age groups: fetal (18-20 weeks gestation), juvenile (5-16 years old), young adults (21-29 years old) and older adults (50-61 years old). We identify 3,523 proteins including 185 ECM proteins and quantify 117 of them. We detect previously unknown proteome and matrisome features during pancreas development and maturation. We also visualize specific ECM proteins of interest using immunofluorescent staining and investigate changes in ECM localization within islet or acinar compartments. This comprehensive proteomics analysis contributes to an improved understanding of the critical roles that ECM plays throughout human pancreas development and maturation.
Highlights
The extracellular matrix (ECM) is unique to each tissue and capable of guiding cell differentiation, migration, morphology, and function
185 proteins were categorized as ECM proteins based on Human Matrisome Database[29,52,53] and 117 were quantifiable (Supplementary Data 4 and 5), which makes it one of the largest datasets of human pancreas matrisome
Discussion dynamic proteome changes are known to occur with aging[2,3,58], a comprehensive analysis of the proteome and matrisome across early and late developmental stages of human pancreatic tissue has not been reported
Summary
The extracellular matrix (ECM) is unique to each tissue and capable of guiding cell differentiation, migration, morphology, and function. We visualize specific ECM proteins of interest using immunofluorescent staining and investigate changes in ECM localization within islet or acinar compartments This comprehensive proteomics analysis contributes to an improved understanding of the critical roles that ECM plays throughout human pancreas development and maturation. The utility of these commercial isobaric tags in large-scale, discovery proteomics studies is often hampered by the limited multiplexing capacity and the high price of the reagent kits To address these limitations, we have developed a cost-effective alternative, based on a set of N,N-dimethyl leucine (DiLeu) isobaric tags that can be synthesized in-house at high yield, with three steps using commercially available reagents, at a fraction of the cost. The multiplexing capacity was increased threefold from 4plex to 12-plex by taking advantage of the mass-defect feature of stable isotopes to enable simultaneous quantification of 12 samples via 12 reporter ions spanning from 115 to 118 m/z48
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