Distribution of Dietary Protein Positively Influences Muscle Protein Synthesis in Healthy
Adults
Wendy Barth, Kerry Peterson, University of Wisconsin-Stout
Abstract
The RDA for protein describes the amount of protein that should be consumed by an individual in a day
to meet the populations needs and to aid in the prevention of protein deficiency (1,2). The consumption
of protein often exceeds the RDA in many countries. Consumption of protein is often skewed rather
than consistent with meals, having the evening meal be more protein dense and morning meal being
typically carbohydrate dense and low in protein (1,2). This study examines the effects of protein
distribution on 24-h skeletal muscle protein synthesis in healthy, adult mean and women. It was
designed with a 7 day crossover feeding and a 30 day washout period. Changes in muscle protein
synthesis were measured in response to protein at breakfast, lunch, and dinner distributed evenly or
skewed (1). During the 24-h periods on days 1 and 7, venous blood samples and vastus lateralis
muscle biopsy samples were obtained during primed constant infusion of phenylalanine. The 24-h
mixed muscle protein fractional synthesis rate was 25% higher in the even than in the skew protein
distribution groups. 24-h muscle protein synthesis is stimulated by the intake of a moderate amount of
protein distributed at each meal in a more effective manner than skewing protein consumption toward
the evening meal (1).
Importance of Protein in Nutrition
• Proteins are found throughout the body, 40% of which is found in skeletal
muscle. The remainder is found in body organs (25%) and mostly in skin
and blood (2)
• Proteins are essential nutritionally because of their constituent amino
acids, which the body must have to synthesize its own variety of
protein and nitrogen containing molecules that make life possible (2)
Break Down of the Amino Acid
An amino acid backbone consists of a central carbon, amino group, a
carboxyl (acid) group, and an R-group (variant). The R-group is what
determines the type of amino acid depending on what side chain is
attached.
Protein Synthesis: Mechanism of Action
The synthesis of proteins takes two steps: transcription and translation.
Transcription takes the information encoded in DNA and encodes it into
mRNA, which heads out of the cell’s nucleus and into the cytoplasm. During
translation, the mRNA works with a ribosome and tRNA to synthesize
proteins (2).
RDA recommendations of Protein
• Adults: 0.8 g of protein per kg body weight per day (g/kg/day)
• Children and adolescence: 1.5-1 g/kg/day
• Women during pregnancy: 0.8- 1.1 g/kg/day
• Woman lactating: 1.3 g/kg/day
Introduction
The current RDA for protein describes the minimum amount of protein that should be consumed in a day
to prevent deficiency (1,2). Muscle protein synthesis over a 24-h period appears to be greater with
consistent consumption of protein at each meal versus unequal intake throughout the day (2).There has
been increasing evidence that demonstrates the increase of dietary protein consumption beyond 0.8 g
protein/kg having beneficial effects on health-related outcomes such as the regulation of muscle mass,
body composition, and function. NHANES provides data that demonstrates that adults in the US often
skew protein intake toward the end of the day at dinner time rather than at breakfast or lunch. It appears
that a meal containing about 30 g of high quality protein, maximizes stimulation of skeletal muscle
protein synthesis in healthy adults (1).
Hypothesis
• An even protein distribution (30 grams at each meal) will result in a greater 24-hour
muscle protein synthetic response compared to an equivalent amount of protein
delivered in a skewed format (10 g at breakfast, 15 g at lunch, 65 g at dinner).
References
1. Mamerow MM, Mettler JA, English KL, Casperson SL, ArentsonLantz E, Sheffield-Moore M, Layman DK,
Paddon-Jones D. Dietary protein distribution positively influences 24-h muscle protein synthesis in healthy
adults. J Nutr 2014;144:876–80.
2. Groff, J., Smith, J., & Gropper, S. (2018) Advanced nutrition and human metabolism. 7th edition. Belmont CA:
Wadsworth Cengage Learning.
3. Katsanos, C. S., Kobayashi, H., Sheffield-Moore, M., Aarsland, A., Wolfe, R. R. (2006). A high proportion of
leucine is required for optimal stimulation of the rate of muscle protein synthesis by essential amino acids in the
elderly. AJP: Endocrinology and Metabolism, 291(2). doi:10.1152/ajpendo.00488.2005
4. Paddon-Jones, D., & Rasmussen, B. B. (2009). Dietary protein recommendations and the prevention of
sarcopenia: Protein, amino acid metabolism and therapy. Current Opinion in Clinical Nutrition and Metabolic
Care, 12(1), 86–90. http://doi.org/10.1097/MCO.0b013e32831cef8b
Methods
Participants: Volunteers were recruited through newspaper advertisements. Medical screenings were conducted
for study eligibility. Screenings included a medical history, blood glucose concentrations, complete blood count,
plasma electrolytes, and liver and renal function tests. Participants were turned away if falling under any of the
following categories: BMI >30 kg/m2, metabolic disease, low hematocrit or hemoglobin, vascular disease,
hypertension, cardiac abnormalities, renal disease recent weight loss or weight gain, participation in a weight
loss/gain exercise program, smoking, and anabolic steroid use (1). The study ended up qualifying eight healthy
individuals; five males and three females between the ages of 25-55 (Table 1). The volunteers in this study were
all physically active but not athletically trained (1).
Test Design: Participants partook in a randomized crossover study. Individuals where given even and skewed
amounts of protein at each meal. All meals given to participants were prepared by ITS-CRC bio-nutrition staff
ensuing control of macronutrients and energy intake in the two diets being tested. Both diets were made to
exceed the RDA to reflect upon current protein consumption in the US (1). The diets being used in this study both
contained ~90 g protein, but were dispersed in different patterns. Diet 1 (EVEN): ~30 g (breakfast), 30 g (lunch),
and 30 g (dinner). Diet 2 (SKEW): ~10 g (breakfast), 15 g (lunch), and 65 g (dinner) (Table 2). Intake of CHO was
consistent; intake of dietary fat was altered to provide the same total daily energy consumption between the two
diets (1).
Time Frame: 7 day crossover feeding with a 30 day washout period (1)
Protocol: During the study, meals were provided at 9:30 am, 1:00pm, and 5:00pm. On days 1 and 7 of each diet,
individuals completed a 24 hr metabolic study, having the second metabolic study on day 7 determine if there was
habituation to the study over the time period it was conducted (1). After their screening, the participants were
asked to continue with their normal diets and avoid any strenuous activity 72 hour prior to each metabolic study.
Participants were admitted to the Institute for Translational Science- Clinical Research at 3 pm the day before the
study. After being provided a standardized evening meal and an overnight fast, they were awaken at 5:30 AM to
have an 18-gauge polyethylene catheter inserted into a forearm vein for blood sampling. Individuals completed a
30 min. period of moderate intensity treadmill walking at 4 PM on days 1 and 7 to reduce any potential
complications associated with reduced physical activity (1).
Measurements for Data Analysis of Hypothesis:
• Phenylalanine enrichment and concentration: An 18-gauge polyethylene catheter inserted into forearm vein
for blood sampling. A second was inserted into other arm for infusion of stable isotope tracer.
• Muscle biopsies: (100-150 mg) taken at 9:00am, 12:30pm, and 24 hours later (9:00am)
• Peripheral venous blood samples: samples taken, once before eating the meal and then every 20 mins. after
for 2-3 hrs.
• Mixed Muscle Protein FSR: measured protein synthesis rate after both EVEN (30g) and SKEW (10g)
breakfasts, and after 24-hr diet, on days 1 and 7 after ingestion.
Results and Outcomes
Dietary Intake
• Table 2 presents a seven-day mean energy and macronutrient intake. The meals on days 1 and 7 that were
provided to participants during the metabolic studies were identical and the total 24-h protein, CHO, and fat
intake were also the same in both the SKEW and EVEN diets. The animal to vegetable protein ration used in
both the SKEW and EVEN diets was ~2:1 for this study (1).
Plasma and muscle enrichments
• The mean plasma phenylalanine enrichments did not differ for either group between the different study days
(day 1 and day 7). Fig. 1 and Table 3 present the plasma and muscle intracellular phenylalanine enrichments
(1).
Mixed muscle protein synthesis
• Breakfast meal: 30 grams vs. 10 grams protein. During day 1 of the metabolic study, muscle protein synthesis
was ~30% higher in the response of the meal containing 30 g protein than the meal with only 10 g protein. Day
7 provided a similar result due to dietary habituation (Fig. 2).
• 24-hr EVEN vs. SKEW protein distribution: Both 24-hr diets contained similar amounts of protein (90-94g).
Day one showed ~25% higher muscle protein synthesis for when dietary protein was distributed evenly by 3
meals/day compared to the SKEW results. Again, day 7 showed similar muscle protein synthesis after 7 days
of habituation to each diet (Fig. 3 and 4).
Overall Results Suggest: participants with the EVEN diet had better muscle protein synthesis when consuming
high-quality protein 3 times a day compared to their SKEWed counterparts. This favors the hypothesis that even
protein distribution results in greater muscle protein synthesis compared to protein consumed in a skewed pattern.
Discussion
Muscle protein synthesis was investigated with the consumption of an evenly dispersed amount of high-quality protein (30g 3x
daily), compared to a skewed delivery (10g,15g, 65g). The muscle protein synthesis after the EVEN diet breakfast, at 30g of
protein, was about 40% higher than the SKEW diet breakfast at 10 g protein. The lunch meals were not directly assessed but
were considered very likely to follow the same pattern favoring the EVEN over the SKEW. Bringing it all together, the total daily
protein consumption in providing the bulk of protein at dinner during the SKEW, failed to make up the difference in 24-hr muscle
protein synthesis proving that the hypothesis was correct in suggesting that an even distribution of high-quality protein being
consumed daily provides an enhanced muscle protein synthesis compared to consuming the same amount of protein daily but
in a skewed format.
- Another study (3) demonstrated the disabled response of muscle protein synthesis after consuming small amounts of amino
acids and how it can be reversed by an intake of extra leucine, an amino acid found in protein. The data emphasized leucine
and its role in any amino acid or protein supplement for reversing the lessened response of muscle protein synthesis in aging
individuals. A higher amount consumed stimulated the initiation of muscle protein synthesis. An evenly dispersed amount went
on to show a greater response in muscle synthesis than the lower or uneven amounts provided to each individual (3).
-Another study (4) demonstrated the aspect of daily protein intake having a direct response with net protein balance in
distribution of protein across three or more meals daily. The study suggested that for a 75 kg person, the RDA states that they
would require 60g protein a day, or if distributed across three meals evenly, 20g protein a day. 20 g serving of most proteins
contain about 5 to 8 g of essential amino acids, which are predominantly responsible for stimulating muscle protein synthesis
suggesting that an intake of this amount of high-quality protein, would be a useful strategy to maintain muscle mass (4).
- The next step would be to further study, for clinical relevance, at what amount synthesis peaks by ingesting higher amounts
of protein, evenly dispersed between meals and the adverse affects on muscle synthesis.
Summary
The ingestion of a moderate amount of high quality protein 3x daily, provides a more effective way of enhancing 24-hr
skeletal muscle protein synthesis than the typical United States diet that skews the majority of protein consumption toward
the evening meal. In conclusion, this study provides results suggesting a moderate, meal-driven approach to daily protein
ingestion having a positive effect on muscle protein synthesis in healthy adults.