Dynamic proteome profiling of individual proteins in human skeletal muscle after a high‐fat diet and resistance exercise

Abstract

It is generally accepted that muscle adaptation to resistance exercise (REX) training is underpinned by contraction‐induced, increased rates of protein synthesis and dietary protein availability. By using dynamic proteome profiling (DPP), we investigated the contribution of both synthesis and breakdown to changes in abundance on a protein‐by‐protein basis in human skeletal muscle. Age‐matched, overweight males consumed 9 d of a high‐fat, low‐carbohydrate diet during which time they either undertook 3 sessions of REX or performed no exercise. Precursor enrichment and the rate of incorporation of deuterium oxide into newly synthesized muscle proteins were determined by mass spectrometry. Ninety proteins were included in the DPP, with 28 proteins exhibiting significant responses to REX. The most common pattern of response was an increase in turnover, followed by an increase in abundance with no detectable increase in protein synthesis. Here, we provide novel evidence that demonstrates that the contribution of synthesis and breakdown to changes in protein abundance induced by REX differ on a protein‐by‐protein basis. We also highlight the importance of the degradation of individual muscle proteins after exercise in human skeletal muscle.—Camera, D. M., Burniston, J. G., Pogson, M. A., Smiles, W. J., Hawley, J. A. Dynamic proteome profiling of individual proteins in human skeletal muscle after a high‐fat diet and resistance exercise. FASEB J. 31, 5478–5494 (2017). http://www.fasebj.org

Reliability of Protein Abundance and Synthesis Measurements in Human Skeletal Muscle

“…Stable isotope labelling of newly synthesised proteins in vivo was achieved by oral consumption of 2 H 2 O over a 14‐day period. Consistent with our previous work, participants consumed 50 mL of 99.8 atom% of 2 H 2 O four times per day. Venous blood was collected bi‐daily, and muscle was collected at baseline (day 0), and after 4, 9, and 14 days of 2 H 2 O consumption.…”

“…Sample size calculations, based on Q 3 biological variation, estimate a required n of 6 (ABD) or 15 (FSR) to detect a within‐subject change of 50% (α = 0.05, 80% power). DPP of human muscle responses to resistance exercise reported changes in FSR that, generally, were of twice the magnitude of changes in ABD. The above observations suggest DPP has an equal ability to detect changes in ABD and FSR in the setting of human exercise physiology.…”

“…Proteins within human muscle exhibit a broad range of different turnover rates and changes to both synthesis and degradation contribute to adaptations in protein abundance . We recently developed the new technique of dynamic proteome profiling (DPP) that can measure both the abundance and synthesis rate of individual proteins in human muscle. DPP combines deuterium oxide ( 2 H 2 O)‐labeling with peptide MS and offers the first insight to dynamic aspects of the human proteome in vivo.…”

“…Body water enrichment of 2 H was measured in plasma samples against external standards by GC–MS . Soluble proteins were extracted from muscle samples as previously described . Tryptic digestion was performed using filter‐aided sample preparation .…”

A single bout of strenuous exercise overcomes lipid‐induced anabolic resistance to protein ingestion in overweight, middle‐aged men

“…Blood glucose concentrations were determined using a YSI (model 2900) Biochemistry Analyzer (Xylem, Hemmant, QLD, Australia). Plasma insulin, TNF-a, and IL-6 were measured utilizing commercially available and customized Milliplex human magnetic bead panels (MilliporeSigma, Burlington, MA, USA) according to the manufacturer's instructions and as previously described in Camera et al (13). Plasma amino acid concentrations were determined by GC-MS (7890A GC/5975C; MSD, 215; Agilent Technologies, Santa Clara, CA, USA) as previously described in Gorissen et al (14).…”