Multiplexed Temporal Quantification of the Exercise-regulated Plasma Peptidome

Area of Science:

• Biochemistry
• Physiology
• Proteomics

Background:

• Exercise offers significant health benefits, potentially mediated by secreted peptide/protein hormones.
• The plasma peptidome, containing low molecular weight peptides, is a rich source of these hormones.
• Understanding exercise-induced changes in the plasma peptidome can identify novel health-related factors.

Purpose of the Study:

• To quantify the effects of intense exercise on the human plasma peptidome.
• To identify novel exercise-regulated secretory factors and understand their mechanisms.
• To validate peptidomics as a method for analyzing physiological perturbations.

Main Methods:

• Developed and optimized a 2D-LC-MS/MS method with HCD and EThcD fragmentation.
• Analyzed endogenous peptides from plasma during exercise and recovery.
• Quantified 5,548 unique peptides, identifying known hormones and novel factors like Transgelin peptides.

Main Results:

• The plasma peptidome showed dynamic, rapid, and reversible changes during exercise.
• Identified known hormones (insulin, glucagon, ghrelin) and predicted bioactive peptides, including Transgelin C-terminal peptides.
• Demonstrated exercise's rapid modulation of hundreds of bioactive peptides via proteases and PTMs.

Conclusions:

• Deep peptidomic analysis reveals exercise rapidly modulates circulating bioactive peptides.
• Transgelin peptides are increased during exercise and influence cell functions in vitro.
• Peptidomics is an effective method for discovering exercise-regulated factors contributing to health benefits.