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. 2011 Oct;111(4):1106-17.
doi: 10.1152/japplphysiol.00631.2011. Epub 2011 Jul 28.

Clustering of GLUT4, TUG, and RUVBL2 protein levels correlate with myosin heavy chain isoform pattern in skeletal muscles, but AS160 and TBC1D1 levels do not

Carlos M Castorena  1 , James G Mackrell, Jonathan S Bogan, Makoto Kanzaki, Gregory D Cartee
Affiliations

Clustering of GLUT4, TUG, and RUVBL2 protein levels correlate with myosin heavy chain isoform pattern in skeletal muscles, but AS160 and TBC1D1 levels do not

Carlos M Castorena et al. J Appl Physiol (1985). 2011 Oct.

Abstract

Skeletal muscle is a heterogeneous tissue. To further elucidate this heterogeneity, we probed relationships between myosin heavy chain (MHC) isoform composition and abundance of GLUT4 and four other proteins that are established or putative GLUT4 regulators [Akt substrate of 160 kDa (AS160), Tre-2/Bub2/Cdc 16-domain member 1 (TBC1D1), Tethering protein containing an UBX-domain for GLUT4 (TUG), and RuvB-like protein two (RUVBL2)] in 12 skeletal muscles or muscle regions from Wistar rats [adductor longus, extensor digitorum longus, epitrochlearis, gastrocnemius (mixed, red, and white), plantaris, soleus, tibialis anterior (red and white), tensor fasciae latae, and white vastus lateralis]. Key results were 1) significant differences found among the muscles (range of muscle expression values) for GLUT4 (2.5-fold), TUG (1.7-fold), RUVBL2 (2.0-fold), and TBC1D1 (2.7-fold), but not AS160; 2) significant positive correlations for pairs of proteins: GLUT4 vs. TUG (R = 0.699), GLUT4 vs. RUVBL2 (R = 0.613), TUG vs. RUVBL2 (R = 0.564), AS160 vs. TBC1D1 (R = 0.293), and AS160 vs. TUG (R = 0.246); 3) significant positive correlations for %MHC-I: GLUT4 (R = 0.460), TUG (R = 0.538), and RUVBL2 (R = 0.511); 4) significant positive correlations for %MHC-IIa: GLUT4 (R = 0.293) and RUVBL2 (R = 0.204); 5) significant negative correlations for %MHC-IIb vs. GLUT4 (R = -0.642), TUG (R = -0.626), and RUVBL2 (R = -0.692); and 6) neither AS160 nor TBC1D1 significantly correlated with MHC isoforms. In 12 rat muscles, GLUT4 abundance tracked with TUG and RUVBL2 and correlated with MHC isoform expression, but was unrelated to AS160 or TBC1D1. Our working hypothesis is that some of the mechanisms that regulate GLUT4 abundance in rat skeletal muscle also influence TUG and RUVBL2 abundance.

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Figures

Fig. 1.
Fig. 1.
Relative abundance (%) of myosin heavy chain (MHC) isoforms in the 12 skeletal muscles or muscle regions. Open bars represent %MHC-I, shaded bars represent %MHC-IIa, solid bars represent %MHC-IIb, and hatched bars represent %MHC-IIx. Values are means ± SE; n = 8 for each muscle or muscle region. AL, adductor longus; EDL, extensor digitorum longus; EPI, epitrochlearis; GASM, mixed gastrocnemius; GASR, red gastrocnemius; GASW, white gastrocnemius; PLAN, plantaris; SOL, soleus; TAR, red tibialis anterior; TAW, white tibialis anterior; TFL, tensor fasciae latae; VLW, white vastus lateralis.
Fig. 2.
Fig. 2.
Relative GLUT4 protein abundance for 12 muscles or muscle regions. *SOL greater than: EDL, EPI, GASM, GASW, TAW, TFL, VLW (P < 0.05); †AL greater than: EPI, GASM, GASW, TAW, TFL, VLW (P < 0.05); #PLAN greater than: EPI, GASW,TAW (P < 0.05); ‡TAR greater than: EPI, GASW, TAW (P < 0.05); ffGASR greater than: EPI, GASW, TAW, VLW; P < 0.05; εEDL greater than: EPI, VLW (P < 0.05). Values are means ± SE; n = 8 for each muscle or muscle region.
Fig. 3.
Fig. 3.
Relative TUG protein abundance for 12 muscles or muscle regions. *SOL and AL were greater than: EPI, EDL, VLW, TAW, GASW (P < 0.05). Values are means ± SE; n = 8 for each muscle or muscle region.
Fig. 4.
Fig. 4.
Relative RUVBL2 protein abundance for 12 muscles or muscle regions. *SOL greater than: GASW, VLW, TAW, EDL (P < 0.05); †AL greater than: VLW, TAW, GASW (P < 0.05); #TFL and PLAN greater than VLW and TAW (P < 0.05); and ‡GASR was greater than the VLW. Values are means ± SE; n = 8 for each muscle or muscle region.
Fig. 5.
Fig. 5.
Relative AS160 protein abundance for 12 muscles or muscle regions. Values are means ± SE; n = 8 for each muscle or muscle region.
Fig. 6.
Fig. 6.
Relative TBC1D1 protein abundance for 12 muscles or muscle regions. *GASW and EDL are greater than EPI and TFL (P < 0.05); †GASM and GASR are greater than TFL (P < 0.05). Values are means ± SE; n = 8 for each muscle or muscle region.
Fig. 7.
Fig. 7.
Correlations between the following pairs of proteins: A: GLUT4 vs. TUG; B: GLUT4 vs. RUVBL2; C: TUG vs. RUVBL2; D: AS160 vs. TBC1D1; E: AS160 vs. TUG. The symbols represent individual data. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 8.
Fig. 8.
Correlations between %MHC-I and the following proteins: A: GLUT4; B: TUG; and C: RUVBL2. The symbols represent individual data. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 9.
Fig. 9.
Correlations between %MHC-IIa and the following proteins: A: GLUT4; B: TUG; and C: RUVBL2. The symbols represent individual data. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 10.
Fig. 10.
Correlations between %MHC-IIb and the following proteins: A: GLUT4; B: TUG; C: and RUVBL2. The symbols represent individual data. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 11.
Fig. 11.
Correlation between %MHC-IIx and the following proteins: A: GLUT4; B: TUG; and C: RUVBL2. The symbols represent individual data. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 12.
Fig. 12.
Correlations between %MHC-I and the following proteins: A: AS160 and B: TBC1D1. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 13.
Fig. 13.
Correlations between %MHC-IIa and the following proteins: A: AS160 and B: TBC1D1. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 14.
Fig. 14.
Correlations between %MHC-IIb and the following proteins: A: AS160 and B: TBC1D1. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
Fig. 15.
Fig. 15.
Correlations between %MHC-IIx and the following proteins: A: AS160 and B: TBC1D1. For correlations using all muscles, n = 96. For correlations excluding the AL and SOL data (w/o AL & SOL), n = 80.
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