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Ask your administrator if you think this is wrong. ======= PRKAA1 ======= == Gene Information == * **<color #00a2e8>Official Symbol</color>**: PRKAA1 * **<color #00a2e8>Official Name</color>**: protein kinase AMP-activated catalytic subunit alpha 1 * **<color #00a2e8>Aliases and Previous Symbols</color>**: N/A * **<color #00a2e8>Entrez ID</color>**: [[https://www.ncbi.nlm.nih.gov/gene/?term=5562|5562]] * **<color #00a2e8>UniProt</color>**: [[https://www.uniprot.org/uniprot/Q13131|Q13131]] * **<color #00a2e8>Interactions</color>**: [[https://thebiogrid.org/search.php?search=PRKAA1&organism=9606|BioGRID]] * **<color #00a2e8>PubMed articles</color>**: [[https://www.ncbi.nlm.nih.gov/pubmed/?term=gene%20PRKAA1|Open PubMed]] * **<color #00a2e8>OMIM</color>**: [[https://omim.org/entry/602739|Open OMIM]] == Function Summary == * **<color #00a2e8>Entrez Summary</color>**: N/A * **<color #00a2e8>UniProt Summary</color>**: Catalytic subunit of AMP-activated protein kinase (AMPK), an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy-consuming processes: inhibits protein, carbohydrate and lipid biosynthesis, as well as cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Regulates lipid synthesis by phosphorylating and inactivating lipid metabolic enzymes such as ACACA, ACACB, GYS1, HMGCR and LIPE; regulates fatty acid and cholesterol synthesis by phosphorylating acetyl-CoA carboxylase (ACACA and ACACB) and hormone-sensitive lipase (LIPE) enzymes, respectively. Regulates insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3. AMPK stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter SLC2A4/GLUT4 to the plasma membrane, possibly by mediating phosphorylation of TBC1D4/AS160. Regulates transcription and chromatin structure by phosphorylating transcription regulators involved in energy metabolism such as CRTC2/TORC2, FOXO3, histone H2B, HDAC5, MEF2C, MLXIPL/ChREBP, EP300, HNF4A, p53/TP53, SREBF1, SREBF2 and PPARGC1A. Acts as a key regulator of glucose homeostasis in liver by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. In response to stress, phosphorylates 'Ser-36' of histone H2B (H2BS36ph), leading to promote transcription. Acts as a key regulator of cell growth and proliferation by phosphorylating TSC2, RPTOR and ATG1/ULK1: in response to nutrient limitation, negatively regulates the mTORC1 complex by phosphorylating RPTOR component of the mTORC1 complex and by phosphorylating and activating TSC2. In response to nutrient limitation, promotes autophagy by phosphorylating and activating ATG1/ULK1. AMPK also acts as a regulator of circadian rhythm by mediating phosphorylation of CRY1, leading to destabilize it. May regulate the Wnt signaling pathway by phosphorylating CTNNB1, leading to stabilize it. Also has tau-protein kinase activity: in response to amyloid beta A4 protein (APP) exposure, activated by CAMKK2, leading to phosphorylation of MAPT/TAU; however the relevance of such data remains unclear in vivo. Also phosphorylates CFTR, EEF2K, KLC1, NOS3 and SLC12A1. {ECO:0000269|PubMed:11518699, ECO:0000269|PubMed:11554766, ECO:0000269|PubMed:12519745, ECO:0000269|PubMed:14651849, ECO:0000269|PubMed:15866171, ECO:0000269|PubMed:17486097, ECO:0000269|PubMed:17711846, ECO:0000269|PubMed:18184930, ECO:0000269|PubMed:18439900, ECO:0000269|PubMed:20074060, ECO:0000269|PubMed:20160076, ECO:0000269|PubMed:21205641}. <button type='primary' size='sm' modal='Pfam_Domains'>Pfam Domains</button> <button type='primary' size='sm' modal='GO_terms'>GO Terms</button> <modal id='Pfam_Domains' size='lg' title='Pfam Domains'> |Pkinase| |Pkinase Tyr| </modal> <modal id='GO_terms' size='lg' title='GO Terms'> |regulation of glucosylceramide biosynthetic process| |negative regulation of glucosylceramide biosynthetic process| |[acetyl-CoA carboxylase] kinase activity| |positive regulation of mitochondrial transcription| |[hydroxymethylglutaryl-CoA reductase (NADPH)] kinase activity| |regulation of mitochondrial transcription| |cold acclimation| |CAMKK-AMPK signaling cascade| |response to camptothecin| |histone serine kinase activity| |negative regulation of ceramide biosynthetic process| |negative regulation of tubulin deacetylation| |calmodulin dependent kinase signaling pathway| |regulation of stress granule assembly| |negative regulation of sphingolipid biosynthetic process| |positive regulation of sterol biosynthetic process| |positive regulation of cholesterol biosynthetic process| |AMP-activated protein kinase activity| |motor behavior| |positive regulation of cholesterol metabolic process| |regulation of tubulin deacetylation| |cAMP-dependent protein kinase activity| |histone-serine phosphorylation| |cellular response to ethanol| |fatty acid homeostasis| |nucleotide-activated protein kinase complex| |cellular response to prostaglandin E stimulus| |regulation of sphingolipid biosynthetic process| |regulation of membrane lipid metabolic process| |regulation of ceramide biosynthetic process| |response to diuretic| |response to caffeine| |positive regulation of steroid biosynthetic process| |tau-protein kinase activity| |cellular response to prostaglandin stimulus| |negative regulation of lipid catabolic process| |response to prostaglandin E| |positive regulation of glycolytic process| |positive regulation of alcohol biosynthetic process| |positive regulation of skeletal muscle tissue development| |regulation of mitochondrial gene expression| |positive regulation of steroid metabolic process| |histone phosphorylation| |response to prostaglandin| |positive regulation of protein targeting to mitochondrion| |positive regulation of peptidyl-lysine acetylation| |negative regulation of insulin receptor signaling pathway| |neuron cellular homeostasis| |energy homeostasis| |negative regulation of cellular response to insulin stimulus| |positive regulation of protein acetylation| |cholesterol biosynthetic process| |cellular response to glucose starvation| |regulation of cholesterol biosynthetic process| |regulation of sterol biosynthetic process| |regulation of protein deacetylation| |secondary alcohol biosynthetic process| |regulation of protein targeting to mitochondrion| |positive regulation of nucleotide metabolic process| |positive regulation of ATP metabolic process| |tau protein binding| |positive regulation of purine nucleotide metabolic process| |sterol biosynthetic process| |negative regulation of lipid biosynthetic process| |negative regulation of TOR signaling| |regulation of skeletal muscle tissue development| |response to cold| |regulation of lipid catabolic process| |cellular response to fatty acid| |response to gamma radiation| |regulation of cholesterol metabolic process| |anion homeostasis| |positive regulation of establishment of protein localization to mitochondrion| |regulation of peptidyl-lysine acetylation| |regulation of insulin receptor signaling pathway| |response to activity| |cellular response to glucose stimulus| |positive regulation of muscle organ development| |positive regulation of striated muscle tissue development| |positive regulation of muscle tissue development| |cellular response to hydrogen peroxide| |cellular response to hexose stimulus| |regulation of protein acetylation| |regulation of establishment of protein localization to mitochondrion| |regulation of cellular response to insulin stimulus| |cellular response to monosaccharide stimulus| |regulation of alcohol biosynthetic process| |fatty acid oxidation| |lipid oxidation| |positive regulation of lipid biosynthetic process| |regulation of glycolytic process| |positive regulation of carbohydrate metabolic process| |cellular response to carbohydrate stimulus| |regulation of protein targeting| |negative regulation of lipid metabolic process| |cellular response to calcium ion| |response to fatty acid| |cellular response to alcohol| |regulation of carbohydrate catabolic process| |cellular glucose homeostasis| |regulation of steroid biosynthetic process| |cellular response to ketone| |response to antineoplastic agent| |regulation of TOR signaling| |protein heterooligomerization| |alcohol biosynthetic process| |response to alkaloid| |glucose metabolic process| |regulation of circadian rhythm| |regulation of steroid metabolic process| |positive regulation of mitochondrion organization| |regulation of purine nucleotide metabolic process| |cholesterol metabolic process| |cellular response to antibiotic| |fatty acid biosynthetic process| |response to hydrogen peroxide| |regulation of nucleotide metabolic process| |positive regulation of autophagy| |regulation of ATP metabolic process| |response to ethanol| |steroid biosynthetic process| |secondary alcohol metabolic process| |cellular response to reactive oxygen species| |regulation of striated muscle tissue development| |lipid homeostasis| |sterol metabolic process| |regulation of muscle tissue development| |regulation of muscle organ development| |regulation of peptidyl-serine phosphorylation| |positive regulation of small molecule metabolic process| |response to glucose| |positive regulation of lipid metabolic process| |response to hexose| |response to UV| |cell cycle arrest| |calcium-mediated signaling| |negative regulation of cellular amide metabolic process| |response to monosaccharide| |response to ionizing radiation| |cellular response to starvation| |response to calcium ion| |response to purine-containing compound| |positive regulation of intracellular protein transport| |activation of MAPK activity| |hexose metabolic process| |regulation of generation of precursor metabolites and energy| |macroautophagy| |response to carbohydrate| |regulation of macroautophagy| |peptidyl-serine phosphorylation| |response to temperature stimulus| |regulation of mitochondrion organization| |regulation of signal transduction by p53 class mediator| |glucose homeostasis| |carbohydrate homeostasis| |organic hydroxy compound biosynthetic process| |monocarboxylic acid biosynthetic process| |regulation of lipid biosynthetic process| |cellular response to hypoxia| |regulation of microtubule cytoskeleton organization| |protein C-terminus binding| |cellular response to metal ion| |response to starvation| |response to ketone| |response to reactive oxygen species| |cellular response to decreased oxygen levels| |peptidyl-serine modification| |monosaccharide metabolic process| |lipid modification| |regulation of carbohydrate metabolic process| |positive regulation of intracellular transport| |cellular response to acid chemical| |cellular response to toxic substance| |cellular response to oxygen levels| |regulation of organelle assembly| |cellular response to inorganic substance| |regulation of microtubule-based process| |regulation of intracellular protein transport| |response to alcohol| |cellular response to nutrient levels| |protein kinase activity| |cellular response to oxidative stress| |steroid metabolic process| |positive regulation of MAP kinase activity| |process utilizing autophagic mechanism| |autophagy| |cellular response to extracellular stimulus| |rhythmic process| |axon| |response to xenobiotic stimulus| |carboxylic acid biosynthetic process| |response to antibiotic| |organic acid biosynthetic process| |response to light stimulus| |negative regulation of catabolic process| |alcohol metabolic process| |apical plasma membrane| |fatty acid metabolic process| |multicellular organismal homeostasis| |positive regulation of cellular protein localization| |activation of protein kinase activity| |regulation of autophagy| |cellular response to external stimulus| |second-messenger-mediated signaling| |positive regulation of protein serine/threonine kinase activity| |anatomical structure homeostasis| |regulation of MAP kinase activity| |response to hypoxia| |response to acid chemical| |regulation of intracellular transport| |response to decreased oxygen levels| |cell-cell signaling by wnt| |Wnt signaling pathway| |protein serine/threonine kinase activity| |histone modification| |positive regulation of cellular catabolic process| |response to metal ion| |neuronal cell body| |covalent chromatin modification| |response to oxygen levels| |response to oxidative stress| |chromatin binding| |regulation of lipid metabolic process| |nuclear speck| |regulation of cellular amide metabolic process| |cellular response to drug| |dendrite| |positive regulation of protein transport| |regulation of small molecule metabolic process| |cell surface receptor signaling pathway involved in cell-cell signaling| |positive regulation of catabolic process| |response to radiation| |regulation of protein complex assembly| |organic hydroxy compound metabolic process| |positive regulation of establishment of protein localization| |carbohydrate metabolic process| |response to nutrient levels| |negative regulation of intracellular signal transduction| |response to toxic substance| |regulation of protein serine/threonine kinase activity| |protein complex oligomerization| |cellular response to lipid| |monocarboxylic acid metabolic process| |response to extracellular stimulus| |positive regulation of protein kinase activity| |response to inorganic substance| |regulation of cellular protein localization| |regulation of cytoskeleton organization| |positive regulation of MAPK cascade| |regulation of vesicle-mediated transport| |behavior| |negative regulation of cell cycle| |positive regulation of kinase activity| |lipid biosynthetic process| |small molecule biosynthetic process| |negative regulation of protein modification process| |cellular response to organonitrogen compound| |cellular response to hormone stimulus| |positive regulation of organelle organization| |positive regulation of transferase activity| |cellular response to nitrogen compound| |chromatin organization| |regulation of protein transport| |regulation of peptide transport| |cellular chemical homeostasis| |regulation of establishment of protein localization| |regulation of MAPK cascade| |ion homeostasis| |regulation of protein kinase activity| |regulation of cellular catabolic process| |response to lipid| |regulation of kinase activity| |peptidyl-amino acid modification| |negative regulation of apoptotic process| |cellular homeostasis| |negative regulation of programmed cell death| |carboxylic acid metabolic process| |response to hormone| |positive regulation of cell population proliferation| |regulation of cellular localization| |response to organic cyclic compound| |cellular lipid metabolic process| |regulation of cellular component biogenesis| |oxidation-reduction process| |protein phosphorylation| |regulation of transferase activity| |positive regulation of transport| |regulation of catabolic process| |negative regulation of cell death| |cell cycle process| |response to organonitrogen compound| |oxoacid metabolic process| |positive regulation of protein phosphorylation| |response to drug| |positive regulation of intracellular signal transduction| |organic acid metabolic process| |regulation of protein localization| |negative regulation of cellular protein metabolic process| |cellular response to oxygen-containing compound| |positive regulation of phosphorylation| |chromosome organization| |response to nitrogen compound| |negative regulation of protein metabolic process| |chemical homeostasis| |cell-cell signaling| |positive regulation of phosphorus metabolic process| |positive regulation of phosphate metabolic process| |response to abiotic stimulus| |regulation of cell cycle| |positive regulation of cellular component organization| |lipid metabolic process| |cellular response to endogenous stimulus| |positive regulation of protein modification process| |negative regulation of signal transduction| |phosphorylation| |regulation of organelle organization| |organic cyclic compound biosynthetic process| |cell cycle| |negative regulation of cell communication| |negative regulation of signaling| |positive regulation of developmental process| |positive regulation of catalytic activity| |regulation of protein phosphorylation| |response to endogenous stimulus| |ATP binding| |negative regulation of cellular biosynthetic process| |regulation of apoptotic process| |positive regulation of transcription, DNA-templated| |negative regulation of biosynthetic process| |response to oxygen-containing compound| |protein-containing complex assembly| |regulation of programmed cell death| |regulation of phosphorylation| |positive regulation of cellular protein metabolic process| |regulation of cell population proliferation| |negative regulation of response to stimulus| |positive regulation of nucleic acid-templated transcription| |positive regulation of RNA biosynthetic process| |homeostatic process| |positive regulation of signal transduction| |regulation of cell death| |intracellular signal transduction| |cellular response to stress| |positive regulation of protein metabolic process| |negative regulation of gene expression| |positive regulation of RNA metabolic process| |positive regulation of multicellular organismal process| |small molecule metabolic process| |positive regulation of molecular function| |regulation of phosphate metabolic process| |regulation of phosphorus metabolic process| |cellular catabolic process| |positive regulation of cell communication| |positive regulation of signaling| |regulation of intracellular signal transduction| |regulation of protein modification process| |protein-containing complex subunit organization| |regulation of transport| |positive regulation of nucleobase-containing compound metabolic process| |positive regulation of macromolecule biosynthetic process| |positive regulation of cellular biosynthetic process| |positive regulation of gene expression| |positive regulation of biosynthetic process| </modal> \\ === CRISPR Data === <button type='primary' size='small' modal='Compound_Hit'>Compound Hit</button> <button type='primary' size='small' modal='Most_Correlated_Genes'>Most Correlated Genes in Chemogenomics</button> <button type='primary' size='small' modal='Essential_Avana'>Tissues where Essential in the Avana Dataset (DepMap 20Q1)</button> <modal id='Compound_Hit' size='lg' title='Compound Hit'> ^Screen^Score^ |[[:results:exp400|Senexin-A 25μM R07 exp400]]|-3.83| |[[:results:exp450|Artemisinin 50μM R08 exp450]]|-3.36| |[[:results:exp85|UM0129480 7μM R02 exp85]]|-3.09| |[[:results:exp447|Amiloride 100μM R08 exp447]]|-3.08| |[[:results:exp445|∆-9-Tetrahydrocannabinol 30μM R08 exp445]]|-3.07| |[[:results:exp275|Citral 75μM R06 exp275]]|-3.02| |[[:results:exp84|UM0125461 0.74μM R02 exp84]]|-3.02| |[[:results:exp499|LY2090314 0.003μM R08 exp499]]|-3| |[[:results:exp500|LY2090314 0.003μM R08 exp500 no dilution day6]]|-3| |[[:results:exp227|Cryptotanshinone 12μM R05 exp227]]|-2.87| |[[:results:exp122|Golgicide-A 4μM R03 exp122]]|-2.87| |[[:results:exp25|Oligomycin-A 2μM R00 exp25]]|-2.81| |[[:results:exp466|Cannabidiol 20μM R08 exp466]]|-2.79| |[[:results:exp495|IWR1 50μM R08 exp495]]|-2.78| |[[:results:exp416|Tubacin 1.6μM R07 exp416]]|-2.72| |[[:results:exp89|Vemurafenib 6.6μM R02 exp89]]|-2.64| |[[:results:exp18|Doxycycline 10μM R00 exp18]]|-2.63| |[[:results:exp317|Geldanamycin 0.015 to 0.05μM on day4 R07 exp317]]|-2.63| |[[:results:exp38|Wortmannin 5μM R00 exp38]]|-2.63| |[[:results:exp472|CI-1040 9.5μM R08 exp472]]|-2.62| |[[:results:exp465|Cannabidiol 13μM R08 exp465]]|-2.58| |[[:results:exp451|Atovaquone 15μM R08 exp451]]|-2.5| |[[:results:exp439|QNZ 0.01μM R08 exp439]]|-2.47| |[[:results:exp231|Epothilone-B 0.0015μM R05 exp231]]|-2.42| |[[:results:exp183|IU1-C 25μM R04 exp183]]|-2.42| |[[:results:exp348|Cyclosporin-A 3μM R07 exp348]]|-2.37| |[[:results:exp334|All-trans-Retinoic-Acid 40μM R07 exp334]]|-2.33| |[[:results:exp66|BI-D1870 3.15μM R02 exp66]]|-2.33| |[[:results:exp517|Quercetin 20μM R08 exp517]]|-2.31| |[[:results:exp350|Deferoxamine 11μM R07 exp350]]|-2.3| |[[:results:exp458|Bisphenol S 100μM R08 exp458]]|-2.28| |[[:results:exp54|Taxol 0.002μM R01 exp54]]|-2.26| |[[:results:exp215|Colchicine 0.009μM R05 exp215]]|-2.22| |[[:results:exp468|CB-5083 0.4μM R08 exp468]]|-2.2| |[[:results:exp35|TRAIL 5ng/ml R00 exp35]]|-2.2| |[[:results:exp448|Ammonium tetrathiomolybdate 10μM R08 exp448]]|-2.16| |[[:results:exp234|Ethanol 0.01 R05 exp234]]|-2.08| |[[:results:exp281|Disulfiram 4.3μM R06 exp281]]|-2.07| |[[:results:exp103|Taxol 0.004μM R03 exp103]]|-2.07| |[[:results:exp492|iCRT14 30μM R08 exp492]]|-2.04| |[[:results:exp365|I-BRD9 4μM R07 exp365]]|-2.01| |[[:results:exp175|3-Bromopyruvate 7μM R04 exp175]]|-1.95| |[[:results:exp502|Milciclib 2μM R08 exp502]]|-1.92| |[[:results:exp37|Wortmannin 0.5μM R00 exp37]]|-1.92| |[[:results:exp442|Ibrutinib 10μM R08 exp442]]|-1.9| |[[:results:exp274|Citral 50μM R06 exp274]]|-1.89| |[[:results:exp532|TIC10 10μM R08 exp532]]|-1.87| |[[:results:exp431|Rotenone 0.07μM R08 exp431]]|-1.83| |[[:results:exp360|Genistein 15μM R07 exp360]]|-1.81| |[[:results:exp311|2-Methoxyestradiol 0.55 to 0.75μM on day4 R07 exp311]]|-1.79| |[[:results:exp359|FK-506 30μM R07 exp359]]|-1.77| |[[:results:exp13|Chloramphenicol 20μM R00 exp13]]|-1.76| |[[:results:exp356|Docosahexaenoic-acid 50μM R07 exp356]]|-1.75| |[[:results:exp120|Dimethyl-Sulfoxide 1pc R03 exp120]]|-1.74| |[[:results:exp455|Benzoate 10000μM R08 exp455]]|-1.71| |[[:results:exp513|ONC212 0.15μM R08 exp513]]|-1.7| |[[:results:exp145|PNU96415E 10μM R03 exp145]]|1.78| |[[:results:exp349|Cytochalasin-B 5μM R07 exp349]]|1.79| |[[:results:exp147|Resveratrol 16μM R03 exp147]]|1.9| |[[:results:exp155|UNC1999 2μM R03 exp155]]|1.95| |[[:results:exp146|Quinacrine 2.5μM R03 exp146]]|2.06| |[[:results:exp510|Nicotine 3000μM R08 exp510]]|2.09| |[[:results:exp508|NN-Dimethylsphingosine 2.5μM R08 exp508]]|2.24| |[[:results:exp222|Betulinic acid 10 to 15μM on day4 R05 exp222]]|2.32| |[[:results:exp414|Tozasertib 0.1μM R07 exp414]]|2.43| |[[:results:exp452|Azithromycin 100μM R08 exp452]]|2.47| |[[:results:exp282|Fluvastatin 2.2μM R06 exp282]]|2.79| |[[:results:exp320|ABT-702 5μM plus CoCl2 18μM R07 exp320]]|2.82| |[[:results:exp301|VER-155008 3.9μM R06 exp301]]|3.17| |[[:results:exp389|PF-06409577 20μM R07 exp389]]|6.45| |[[:results:exp264|Arsenate 40μM R06 exp264]]|7.41| </modal> <modal id='Most_Correlated_Genes' size='lg' title='Most Correlated Genes in Chemogenomics'> ^Gene^Correlation^ |[[:human genes:p:prkag1|PRKAG1]]|0.709| |[[:human genes:p:prkab1|PRKAB1]]|0.659| |[[:human genes:s:strada|STRADA]]|0.62| |[[:human genes:s:stk11|STK11]]|0.537| |[[:human genes:n:nprl3|NPRL3]]|0.411| |[[:human genes:s:szt2|SZT2]]|0.402| </modal> <modal id='Essential_Avana' size='lg' title='Tissues where Essential in the Avana Dataset (DepMap 20Q1)'> Global Fraction of Cell Lines Where Essential: 1/726 ^Tissue^Fraction Of Cell Lines Where Essential^ |1290807.0|0/1| |909776.0|0/1| |bile duct|0/28| |blood|0/28| |bone|0/25| |breast|0/33| |central nervous system|0/56| |cervix|0/4| |colorectal|0/17| |esophagus|0/13| |fibroblast|0/1| |gastric|0/15| |kidney|0/21| |liver|0/20| |lung|0/75| |lymphocyte|0/14| |ovary|0/26| |pancreas|0/24| |peripheral nervous system|0/16| |plasma cell|1/15| |prostate|0/1| |skin|0/24| |soft tissue|0/7| |thyroid|0/2| |upper aerodigestive|0/22| |urinary tract|0/29| |uterus|0/5| </modal> == Essentiality in NALM6 == * **<color #00a2e8>Essentiality Rank</color>**: 8526 * **<color #00a2e8>Expression level (log2 read counts)</color>**: 6.51 <button type='primary' size='small' modal='Dist_expr'>Expression Distribution</button> <modal id='Dist_expr' size='lg' title='PRKAA1 Expression in NALM6 Cells: 6.51'> {{:chemogenomics:nalm6 dist.png?nolink |}} </modal> Last modified: 2025/12/10 20:19by 127.0.0.1