LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle

The influx of essential amino acids into skeletal muscle is primarily mediated by the large neutral amino acid transporter 1 (LAT1), which is dependent on the glutamine gradient generated by the sodium-dependent neutral amino acid transporter 2 (SNAT2). The protein expression and membrane localizati...

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Autores principales: Michael Mazzulla, Nathan Hodson, Matthew Lees, Paula J. Scaife, Kenneth Smith, Philip J. Atherton, Dinesh Kumbhare, Daniel R. Moore
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Publicado: MDPI AG 2021
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spelling oai:doaj.org-article:ff70c40e1499446093c07c9ae33d01142021-11-25T18:35:08ZLAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle10.3390/nu131139062072-6643https://doaj.org/article/ff70c40e1499446093c07c9ae33d01142021-10-01T00:00:00Zhttps://www.mdpi.com/2072-6643/13/11/3906https://doaj.org/toc/2072-6643The influx of essential amino acids into skeletal muscle is primarily mediated by the large neutral amino acid transporter 1 (LAT1), which is dependent on the glutamine gradient generated by the sodium-dependent neutral amino acid transporter 2 (SNAT2). The protein expression and membrane localization of LAT1 may be influenced by amino acid ingestion and/or resistance exercise, although its acute influence on dietary amino acid incorporation into skeletal muscle protein has not been investigated. In a group design, healthy males consumed a mixed carbohydrate (0.75 g·kg<sup>−1</sup>) crystalline amino acid (0.25 g·kg<sup>−1</sup>) beverage enriched to 25% and 30% with LAT1 substrates L-[1-<sup>13</sup>C]leucine (LEU) and L-[<i>ring</i>-<sup>2</sup>H<sub>5</sub>]phenylalanine (PHE), respectively, at rest (FED: <i>n</i> = 7, 23 ± 5 y, 77 ± 4 kg) or after a bout of resistance exercise (EXFED: <i>n</i> = 7, 22 ± 2 y, 78 ± 11 kg). Postprandial muscle biopsies were collected at 0, 120, and 300 min to measure transporter protein expression (immunoblot), LAT1 membrane localization (immunofluorescence), and dietary amino acid incorporation into myofibrillar protein (ΔLEU and ΔPHE). Basal LAT1 and SNAT2 protein contents were correlated with each other (<i>r</i> = 0.55, <i>p</i> = 0.04) but their expression did not change across time in FED or EXFED (all, <i>p</i> > 0.05). Membrane localization of LAT1 did not change across time in FED or EXFED whether measured as outer 1.5 µm intensity or membrane-to-fiber ratio (all, <i>p</i> > 0.05). Basal SNAT2 protein expression was not correlated with ΔLEU or ΔPHE (all, <i>p</i> ≥ 0.05) whereas basal LAT1 expression was negatively correlated with ΔPHE in FED (<i>r</i> = −0.76, <i>p</i> = 0.04) and EXFED (<i>r</i> = −0.81, <i>p</i> = 0.03) but not ΔLEU (<i>p</i> > 0.05). Basal LAT1 membrane localization was not correlated with ΔLEU or ΔPHE (all, <i>p</i> > 0.05). Our results suggest that LAT1/SNAT2 protein expression and LAT1 membrane localization are not influenced by acute anabolic stimuli and do not positively influence the incorporation of dietary amino acids for de novo myofibrillar protein synthesis in healthy young males.Michael MazzullaNathan HodsonMatthew LeesPaula J. ScaifeKenneth SmithPhilip J. AthertonDinesh KumbhareDaniel R. MooreMDPI AGarticledietary proteinresistance exerciseproteinskeletal muscleamino acid transportersNutrition. Foods and food supplyTX341-641ENNutrients, Vol 13, Iss 3906, p 3906 (2021)
institution DOAJ
collection DOAJ
language EN
topic dietary protein
resistance exercise
protein
skeletal muscle
amino acid transporters
Nutrition. Foods and food supply
TX341-641
spellingShingle dietary protein
resistance exercise
protein
skeletal muscle
amino acid transporters
Nutrition. Foods and food supply
TX341-641
Michael Mazzulla
Nathan Hodson
Matthew Lees
Paula J. Scaife
Kenneth Smith
Philip J. Atherton
Dinesh Kumbhare
Daniel R. Moore
LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle
description The influx of essential amino acids into skeletal muscle is primarily mediated by the large neutral amino acid transporter 1 (LAT1), which is dependent on the glutamine gradient generated by the sodium-dependent neutral amino acid transporter 2 (SNAT2). The protein expression and membrane localization of LAT1 may be influenced by amino acid ingestion and/or resistance exercise, although its acute influence on dietary amino acid incorporation into skeletal muscle protein has not been investigated. In a group design, healthy males consumed a mixed carbohydrate (0.75 g·kg<sup>−1</sup>) crystalline amino acid (0.25 g·kg<sup>−1</sup>) beverage enriched to 25% and 30% with LAT1 substrates L-[1-<sup>13</sup>C]leucine (LEU) and L-[<i>ring</i>-<sup>2</sup>H<sub>5</sub>]phenylalanine (PHE), respectively, at rest (FED: <i>n</i> = 7, 23 ± 5 y, 77 ± 4 kg) or after a bout of resistance exercise (EXFED: <i>n</i> = 7, 22 ± 2 y, 78 ± 11 kg). Postprandial muscle biopsies were collected at 0, 120, and 300 min to measure transporter protein expression (immunoblot), LAT1 membrane localization (immunofluorescence), and dietary amino acid incorporation into myofibrillar protein (ΔLEU and ΔPHE). Basal LAT1 and SNAT2 protein contents were correlated with each other (<i>r</i> = 0.55, <i>p</i> = 0.04) but their expression did not change across time in FED or EXFED (all, <i>p</i> > 0.05). Membrane localization of LAT1 did not change across time in FED or EXFED whether measured as outer 1.5 µm intensity or membrane-to-fiber ratio (all, <i>p</i> > 0.05). Basal SNAT2 protein expression was not correlated with ΔLEU or ΔPHE (all, <i>p</i> ≥ 0.05) whereas basal LAT1 expression was negatively correlated with ΔPHE in FED (<i>r</i> = −0.76, <i>p</i> = 0.04) and EXFED (<i>r</i> = −0.81, <i>p</i> = 0.03) but not ΔLEU (<i>p</i> > 0.05). Basal LAT1 membrane localization was not correlated with ΔLEU or ΔPHE (all, <i>p</i> > 0.05). Our results suggest that LAT1/SNAT2 protein expression and LAT1 membrane localization are not influenced by acute anabolic stimuli and do not positively influence the incorporation of dietary amino acids for de novo myofibrillar protein synthesis in healthy young males.
format article
author Michael Mazzulla
Nathan Hodson
Matthew Lees
Paula J. Scaife
Kenneth Smith
Philip J. Atherton
Dinesh Kumbhare
Daniel R. Moore
author_facet Michael Mazzulla
Nathan Hodson
Matthew Lees
Paula J. Scaife
Kenneth Smith
Philip J. Atherton
Dinesh Kumbhare
Daniel R. Moore
author_sort Michael Mazzulla
title LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle
title_short LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle
title_full LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle
title_fullStr LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle
title_full_unstemmed LAT1 and SNAT2 Protein Expression and Membrane Localization of LAT1 Are Not Acutely Altered by Dietary Amino Acids or Resistance Exercise Nor Positively Associated with Leucine or Phenylalanine Incorporation in Human Skeletal Muscle
title_sort lat1 and snat2 protein expression and membrane localization of lat1 are not acutely altered by dietary amino acids or resistance exercise nor positively associated with leucine or phenylalanine incorporation in human skeletal muscle
publisher MDPI AG
publishDate 2021
url https://doaj.org/article/ff70c40e1499446093c07c9ae33d0114
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