# Ampk alpha2 T172 activation dictates exercise performance and energy transduction in skeletal muscle **Authors:** Ryan N. Montalvo, Xiaolu Li, Gina M. Many, Tyler J. Sagendorf, Qing Yu, Wenqing Shen, Nishikant Wase, A. Robert Burgardt, Tong Zhang, Marina A. Gritsenko, Matthew J. Gaffrey, Hemangi Bhonsle, Yuntian Guan, Xuansong Mao, Mei Zhang, Wei-Jun Qian, Zhen Yan PMC · DOI: 10.1126/sciadv.aeb3338 · Science Advances · 2026-02-25 ## TL;DR This study shows that AMPKα2 T172 activation is crucial for muscle energy metabolism, exercise performance, and may be a target for treating type 2 diabetes. ## Contribution The study introduces a novel nonactivatable Ampkα2 KI mouse model to study AMPK function without disrupting protein stoichiometry. ## Key findings - Ampkα2 T172A KI mice show impaired endurance and mitochondrial function in skeletal muscle. - Multiomics analysis reveals the role of Ampkα2 T172 in glycolytic and oxidative metabolism. - Proteomic changes in KI mice overlap with those in type 2 diabetes patients. ## Abstract Adenosine 5′-monophosphate–activated protein kinase (AMPK) is an energetic sensor for metabolic regulation and integration. Here, we used CRISPR-Cas9 to generate nonactivatable Ampkα knock-in (KI) mice with mutation of threonine-172 phosphorylation site to alanine (T172A), circumventing the limitations of previous genetic interventions that disrupt the protein stoichiometry. KI mice of Ampkα2, but not Ampkα1, demonstrated phenotypic changes with increased fat-to-lean mass, impaired endurance exercise capacity, and diminished mitochondrial maximal respiration and conductance in skeletal muscle. Integrated temporal multiomics analysis (proteomics/phosphoproteomics/metabolomics) in skeletal muscle at rest and during exercise establishes a pleiotropic yet imperative role of Ampkα2 T172 activation for glycolytic and oxidative metabolism, mitochondrial respiration, and contractile function. There is a substantial overlap of skeletal muscle proteomic changes in Ampkα2 T172A KI mice with that of patients with type 2 diabetes. Our findings suggest that Ampkα2 T172 activation is critical for exercise performance and energy transduction in skeletal muscle and may serve as a therapeutic target for type 2 diabetes. Integrative phenotypic and multiomic analysis reveals distinct AMPKα2 T172 regulatory clusters driving muscle energy metabolism. ## Linked entities - **Genes:** PRKAA1 (protein kinase AMP-activated catalytic subunit alpha 1) [NCBI Gene 5562], PRKAA2 (protein kinase AMP-activated catalytic subunit alpha 2) [NCBI Gene 5563] - **Proteins:** PRKAA1 (protein kinase AMP-activated catalytic subunit alpha 1), PRKAA2 (protein kinase AMP-activated catalytic subunit alpha 2) - **Diseases:** type 2 diabetes (MONDO:0005148) - **Species:** Mus musculus (taxon 10090) ## Full-text entities - **Genes:** Mtor (mechanistic target of rapamycin kinase) [NCBI Gene 56717] {aka 2610315D21Rik, FRAP, FRAP2, Frap1, RAFT1, RAPT1}, Ndufa8 (NADH:ubiquinone oxidoreductase subunit A8) [NCBI Gene 68375] {aka 0610033L03Rik, CI-19kD, CI-PGIV}, Dst (dystonin) [NCBI Gene 13518] {aka 2310001O04Rik, A830042E19Rik, BP230, BPAG1-n, Bpag, Bpag1}, Eno3 (enolase 3, beta muscle) [NCBI Gene 13808] {aka Eno-3, MSE}, Slc25a12 (solute carrier family 25 (mitochondrial carrier, Aralar), member 12) [NCBI Gene 78830] {aka 2610002D09Rik, B230107K20Rik}, Mpc1 (mitochondrial pyruvate carrier 1) [NCBI Gene 55951] {aka 0610006G08Rik, 3830411I18Rik, Brp44l}, Etfa (electron transferring flavoprotein, alpha polypeptide) [NCBI Gene 110842] {aka 2010200I21Rik, D9Ertd394e}, Ulk1 (unc-51 like kinase 1) [NCBI Gene 22241] {aka Unc51.1, mKIAA0722}, Dlat (dihydrolipoamide S-acetyltransferase) [NCBI Gene 235339] {aka 6332404G05Rik, DLTA, PDC-E2}, Idh3a (isocitrate dehydrogenase 3 (NAD+) alpha) [NCBI Gene 67834] {aka 1110003P10Rik, 1500012E04Rik}, Hadh (hydroxyacyl-Coenzyme A dehydrogenase) [NCBI Gene 15107] {aka HCDH, Hadhsc, Schad}, Prkaa1 (protein kinase, AMP-activated, alpha 1 catalytic subunit) [NCBI Gene 105787] {aka AMPKalpha1, C130083N04Rik}, Sdha (succinate dehydrogenase complex, subunit A, flavoprotein (Fp)) [NCBI Gene 66945] {aka 1500032O14Rik, 2310034D06Rik, 4921513A11, FP, SDH2, SDHF}, Ndufv1 (NADH:ubiquinone oxidoreductase core subunit V1) [NCBI Gene 17995] {aka CI-51kD}, Acc (anterior capsular cataract) [NCBI Gene 104371], Bcl2a1a (B cell leukemia/lymphoma 2 related protein A1a) [NCBI Gene 12044] {aka A1, Bcl2a1, Bfl-1, Hbpa1}, Crtc2 (CREB regulated transcription coactivator 2) [NCBI Gene 74343] {aka 4632407F12Rik, Torc2}, Cycs (cytochrome c, somatic) [NCBI Gene 13063], Ppp1r12b (protein phosphatase 1, regulatory subunit 12B) [NCBI Gene 329251] {aka 1810037O03Rik, 9530009M10Rik, Mypt2}, Slc25a4 (solute carrier family 25 (mitochondrial carrier, adenine nucleotide translocator), member 4) [NCBI Gene 11739] {aka Ant1, mANC1}, Tbc1d1 (TBC1 domain family, member 1) [NCBI Gene 57915] {aka 1110062G02Rik, Nob1, Nobq1, Tbc1, mKIAA1108}, Szrd1 (SUZ RNA binding domain containing 1) [NCBI Gene 213491] {aka 1110022I03, D4Ertd22e}, Capn3 (calpain 3) [NCBI Gene 12335] {aka Capa-3, Capa3, Lp82, p94}, Fnip1 (folliculin interacting protein 1) [NCBI Gene 216742] {aka A730024A03Rik}, Hsdl2 (hydroxysteroid dehydrogenase like 2) [NCBI Gene 72479] {aka 2610207I16Rik}, Ppp1r3a (protein phosphatase 1, regulatory subunit 3A) [NCBI Gene 140491] {aka GM, RG1, RGL}, Pdpr (pyruvate dehydrogenase phosphatase regulatory subunit) [NCBI Gene 319518] {aka 4930402E16Rik, mKIAA1990}, St3gal5 (ST3 beta-galactoside alpha-2,3-sialyltransferase 5) [NCBI Gene 20454] {aka 3S-T, Siat9, [a]2}, Slc25a5 (solute carrier family 25 (mitochondrial carrier, adenine nucleotide translocator), member 5) [NCBI Gene 11740] {aka Ant2}, Hsd17b10 (hydroxysteroid (17-beta) dehydrogenase 10) [NCBI Gene 15108] {aka 17bHSD10, Ads9, ERAB, Hadh2, MHBD}, Svil (supervillin) [NCBI Gene 225115] {aka B430302E16Rik}, Ndufab1 (NADH:ubiquinone oxidoreductase subunit AB1) [NCBI Gene 70316] {aka 2210401F17Rik, 2310039H15Rik, 2610003B19Rik, 8kDa, 9130423F15Rik, ACP}, Etfb (electron transferring flavoprotein, beta polypeptide) [NCBI Gene 110826] {aka 0610009I16Rik, 2810441H06Rik}, Tim (translocation induced circling mutation) [NCBI Gene 107698], Ndufa6 (NADH:ubiquinone oxidoreductase subunit A6) [NCBI Gene 67130] {aka 14kDa, 2700038D15Rik, B230217P19Rik}, Mpc2 (mitochondrial pyruvate carrier 2) [NCBI Gene 70456] {aka 0610006C01Rik, 2010002I07Rik, 2610205H19Rik, Brp44, ESTM43}, Jph1 (junctophilin 1) [NCBI Gene 57339] {aka JP-1, Jp1}, Ttn (titin) [NCBI Gene 22138] {aka 1100001C23Rik, 2310036G12Rik, 2310057K23Rik, 2310074I15Rik, D330041I19Rik, D830007G01Rik}, Uqcr10 (ubiquinol-cytochrome c reductase, complex III subunit X) [NCBI Gene 66152] {aka 1110020P15Rik, Ucrc}, Ckm (creatine kinase, muscle) [NCBI Gene 12715] {aka CPK-M, Ckmm, M-CK, MCK}, Hspe1 (heat shock protein 1 (chaperonin 10)) [NCBI Gene 15528] {aka 10kDa, Hsp10, mt-cpn10}, Nnt (nicotinamide nucleotide transhydrogenase) [NCBI Gene 18115] {aka 4930423F13Rik}, Calm2 (calmodulin 2) [NCBI Gene 12314] {aka 1500001E21Rik, Cam2, CamC}, Pdp1 (pyruvate dehydrogenase phosphatase catalytic subunit 1) [NCBI Gene 381511] {aka Gm1024, PDPC 1, Ppm2c}, Pkn2 (protein kinase N2) [NCBI Gene 109333] {aka 6030436C20Rik, PRK2, Prkcl2, Stk7}, Nos1 (nitric oxide synthase 1, neuronal) [NCBI Gene 18125] {aka 2310005C01Rik, N-NOS, NC-NOS, NO, NOS, NOS-I}, Synpo (synaptopodin) [NCBI Gene 104027] {aka 9030217H17Rik, 9130229N11, 9330140I15Rik}, Pdk2 (pyruvate dehydrogenase kinase, isoenzyme 2) [NCBI Gene 18604], Ckmt2 (creatine kinase, mitochondrial 2) [NCBI Gene 76722] {aka 2300008A19Rik, S-MtCK, ScCKmit, mib-CK}, Pdhx (pyruvate dehydrogenase complex, component X) [NCBI Gene 27402] {aka E3bp, Pdx1}, Alb (albumin) [NCBI Gene 11657] {aka Alb-1, Alb1, BCL001, BCL002, BPL001}, Acadm (acyl-Coenzyme A dehydrogenase, medium chain) [NCBI Gene 11364] {aka MCAD}, Prkaa2 (protein kinase, AMP-activated, alpha 2 catalytic subunit) [NCBI Gene 108079] {aka 2310008I11Rik, A830082D05, AMPKalpha2}, Pdk4 (pyruvate dehydrogenase kinase, isoenzyme 4) [NCBI Gene 27273], Cox4i1 (cytochrome c oxidase subunit 4I1) [NCBI Gene 12857] {aka COX, COX IV-1, COXIV, Cox4, Cox4a, IV-1}, Mylk4 (myosin light chain kinase family, member 4) [NCBI Gene 238564] {aka EG238564}, Maf1 (MAF1 homolog, negative regulator of RNA polymerase III) [NCBI Gene 68877] {aka 1110068E11Rik}, Slc25a3 (solute carrier family 25 (mitochondrial carrier, phosphate carrier), member 3) [NCBI Gene 18674] {aka 5730556H19Rik, PTP, Phc}, Acly (ATP citrate lyase) [NCBI Gene 104112] {aka A730098H14Rik}, Jph2 (junctophilin 2) [NCBI Gene 59091] {aka 1110002E14Rik, JP-2, Jp2} - **Diseases:** diabetes (MESH:D003920), shock (MESH:D012769), metabolic disease (MESH:D008659), metabolic dysregulation (MESH:D021081), fatigue (MESH:D005221), insulin-resistant (MESH:D007333), CS (MESH:D006223), chronic (MESH:D002908), impairments in muscle function (MESH:D009135), EXH (MESH:D006359), T2D (MESH:D003924) - **Chemicals:** ADP (MESH:D000244), G1P (MESH:C031590), deoxyglucose (MESH:D003847), acetonitrile (MESH:C032159), methylmalonic acid (MESH:D008764), canola oil (MESH:D000074262), polyacrylamide (MESH:C016679), TCA (MESH:D014233), lactate (MESH:D019344), nitrogen (MESH:D009584), bromophenol blue (MESH:D001978), EDTA (MESH:D004492), phosphate (MESH:D010710), acid (MESH:D000143), formic acid (MESH:C030544), O2 (MESH:D010100), leupeptin (MESH:C032854), pyruvate (MESH:D019289), NaCl (MESH:D012965), methanol (MESH:D000432), malate (MESH:C030298), Blood glucose (MESH:D001786), DTT (MESH:D004229), SDS (MESH:D012967), adenine (MESH:D000225), l-kynurenine (MESH:D007737), isoflurane (MESH:D007530), Glycogen (MESH:D006003), peptide (MESH:D010455), propionic acid (MESH:C029658), iodoacetamide (MESH:D007460), NaF (MESH:D012969), water (MESH:D014867), 2-mercaptoethanol (MESH:D008623), oxidized glutathione (MESH:D019803), fatty acid (MESH:D005227), carbohydrate (MESH:D002241), creatine (MESH:D003401), phosphocreatine (MESH:D010725), hydroxylamine (MESH:D019811), phenylmethylsulfonyl fluoride (MESH:D010664), l-aspartic acid (MESH:D001224), urea (MESH:D014508), silica (MESH:D012822), Hepes (MESH:D006531), l-carnitine (MESH:D002331), glycerol (MESH:D005990), aluminum (MESH:D000535), JO2 (-), D (MESH:D003903), trehalose (MESH:D014199), Glucose (MESH:D005947), FADH2 (MESH:C058805), FA (MESH:D005492), calcium (MESH:D002118), NAD+ (MESH:D009243), KCl (MESH:D011189), sucrose (MESH:D013395), chloroform (MESH:D002725), agarose (MESH:D012685) - **Species:** Mus musculus (house mouse, species) [taxon 10090], Homo sapiens (human, species) [taxon 9606] - **Mutations:** C to G, ACT to GCT, R to T, E to H, Aspartate 157 Alanine, T712A, AGC of 1, Lysine 45 Arginine, T172, A to D, C to F, M to P, U to Y, T712, I to L, (F) at 10 ## Full text _Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12935046/full.md ## Figures 5 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12935046/full.md ## References 87 references — full list in the complete paper: https://tomesphere.com/paper/PMC12935046/full.md --- Source: https://tomesphere.com/paper/PMC12935046