Product: mTOR Antibody
Catalog: AF6308
Description: Rabbit polyclonal antibody to mTOR
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat, Fish
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 250-289 kDa; 289kD(Calculated).
Uniprot: P42345
RRID: AB_2835169

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat,Fish
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
mTOR Antibody detects endogenous levels of total mTOR.
RRID:
AB_2835169
Cite Format: Affinity Biosciences Cat# AF6308, RRID:AB_2835169.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1); FK506 binding protein 12 rapamycin associated protein 1; FK506 binding protein 12 rapamycin associated protein 2; FK506 binding protein 12 rapamycin complex associated protein 1; FK506-binding protein 12-rapamycin complex-associated protein 1; FKBP rapamycin associated protein; FKBP12 rapamycin complex associated protein; FKBP12-rapamycin complex-associated protein 1; FKBP12-rapamycin complex-associated protein; FLJ44809; FRAP; FRAP1; FRAP2; Mammalian target of rapamycin; Mechanistic target of rapamycin; mTOR; MTOR_HUMAN; OTTHUMP00000001983; RAFT1; Rapamycin and FKBP12 target 1; Rapamycin associated protein FRAP2; Rapamycin target protein 1; Rapamycin target protein; RAPT1; Serine/threonine-protein kinase mTOR;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P42345 MTOR_HUMAN:

Expressed in numerous tissues, with highest levels in testis.

Description:
an atypical kinase belonging to the PIKK family of kinases. Controls cell growth through protein synthesis regulation. Downstream of PI3K/Akt pathway and required for cell survival. Acts as the target for the cell-cycle arrest and immunosuppressive effects of the FKBP12-rapamycin complex.
Sequence:
MLGTGPAAATTAATTSSNVSVLQQFASGLKSRNEETRAKAAKELQHYVTMELREMSQEESTRFYDQLNHHIFELVSSSDANERKGGILAIASLIGVEGGNATRIGRFANYLRNLLPSNDPVVMEMASKAIGRLAMAGDTFTAEYVEFEVKRALEWLGADRNEGRRHAAVLVLRELAISVPTFFFQQVQPFFDNIFVAVWDPKQAIREGAVAALRACLILTTQREPKEMQKPQWYRHTFEEAEKGFDETLAKEKGMNRDDRIHGALLILNELVRISSMEGERLREEMEEITQQQLVHDKYCKDLMGFGTKPRHITPFTSFQAVQPQQSNALVGLLGYSSHQGLMGFGTSPSPAKSTLVESRCCRDLMEEKFDQVCQWVLKCRNSKNSLIQMTILNLLPRLAAFRPSAFTDTQYLQDTMNHVLSCVKKEKERTAAFQALGLLSVAVRSEFKVYLPRVLDIIRAALPPKDFAHKRQKAMQVDATVFTCISMLARAMGPGIQQDIKELLEPMLAVGLSPALTAVLYDLSRQIPQLKKDIQDGLLKMLSLVLMHKPLRHPGMPKGLAHQLASPGLTTLPEASDVGSITLALRTLGSFEFEGHSLTQFVRHCADHFLNSEHKEIRMEAARTCSRLLTPSIHLISGHAHVVSQTAVQVVADVLSKLLVVGITDPDPDIRYCVLASLDERFDAHLAQAENLQALFVALNDQVFEIRELAICTVGRLSSMNPAFVMPFLRKMLIQILTELEHSGIGRIKEQSARMLGHLVSNAPRLIRPYMEPILKALILKLKDPDPDPNPGVINNVLATIGELAQVSGLEMRKWVDELFIIIMDMLQDSSLLAKRQVALWTLGQLVASTGYVVEPYRKYPTLLEVLLNFLKTEQNQGTRREAIRVLGLLGALDPYKHKVNIGMIDQSRDASAVSLSESKSSQDSSDYSTSEMLVNMGNLPLDEFYPAVSMVALMRIFRDQSLSHHHTMVVQAITFIFKSLGLKCVQFLPQVMPTFLNVIRVCDGAIREFLFQQLGMLVSFVKSHIRPYMDEIVTLMREFWVMNTSIQSTIILLIEQIVVALGGEFKLYLPQLIPHMLRVFMHDNSPGRIVSIKLLAAIQLFGANLDDYLHLLLPPIVKLFDAPEAPLPSRKAALETVDRLTESLDFTDYASRIIHPIVRTLDQSPELRSTAMDTLSSLVFQLGKKYQIFIPMVNKVLVRHRINHQRYDVLICRIVKGYTLADEEEDPLIYQHRMLRSGQGDALASGPVETGPMKKLHVSTINLQKAWGAARRVSKDDWLEWLRRLSLELLKDSSSPSLRSCWALAQAYNPMARDLFNAAFVSCWSELNEDQQDELIRSIELALTSQDIAEVTQTLLNLAEFMEHSDKGPLPLRDDNGIVLLGERAAKCRAYAKALHYKELEFQKGPTPAILESLISINNKLQQPEAAAGVLEYAMKHFGELEIQATWYEKLHEWEDALVAYDKKMDTNKDDPELMLGRMRCLEALGEWGQLHQQCCEKWTLVNDETQAKMARMAAAAAWGLGQWDSMEEYTCMIPRDTHDGAFYRAVLALHQDLFSLAQQCIDKARDLLDAELTAMAGESYSRAYGAMVSCHMLSELEEVIQYKLVPERREIIRQIWWERLQGCQRIVEDWQKILMVRSLVVSPHEDMRTWLKYASLCGKSGRLALAHKTLVLLLGVDPSRQLDHPLPTVHPQVTYAYMKNMWKSARKIDAFQHMQHFVQTMQQQAQHAIATEDQQHKQELHKLMARCFLKLGEWQLNLQGINESTIPKVLQYYSAATEHDRSWYKAWHAWAVMNFEAVLHYKHQNQARDEKKKLRHASGANITNATTAATTAATATTTASTEGSNSESEAESTENSPTPSPLQKKVTEDLSKTLLMYTVPAVQGFFRSISLSRGNNLQDTLRVLTLWFDYGHWPDVNEALVEGVKAIQIDTWLQVIPQLIARIDTPRPLVGRLIHQLLTDIGRYHPQALIYPLTVASKSTTTARHNAANKILKNMCEHSNTLVQQAMMVSEELIRVAILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQLPQLTSLELQYVSPKLLMCRDLELAVPGTYDPNQPIIRIQSIAPSLQVITSKQRPRKLTLMGSNGHEFVFLLKGHEDLRQDERVMQLFGLVNTLLANDPTSLRKNLSIQRYAVIPLSTNSGLIGWVPHCDTLHALIRDYREKKKILLNIEHRIMLRMAPDYDHLTLMQKVEVFEHAVNNTAGDDLAKLLWLKSPSSEVWFDRRTNYTRSLAVMSMVGYILGLGDRHPSNLMLDRLSGKILHIDFGDCFEVAMTREKFPEKIPFRLTRMLTNAMEVTGLDGNYRITCHTVMEVLREHKDSVMAVLEAFVYDPLLNWRLMDTNTKGNKRSRTRTDSYSAGQSVEILDGVELGEPAHKKTGTTVPESIHSFIGDGLVKPEALNKKAIQIINRVRDKLTGRDFSHDDTLDVPTQVELLIKQATSHENLCQCYIGWCPFW

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Chicken
100
Rabbit
100
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P42345 As Substrate

Site PTM Type Enzyme
M1 Acetylation
K42 Ubiquitination
K84 Ubiquitination
S92 Phosphorylation
T102 Phosphorylation
Y110 Phosphorylation
K128 Ubiquitination
K230 Ubiquitination
K243 Ubiquitination
K251 Ubiquitination
K298 Ubiquitination
K301 Ubiquitination
T308 Phosphorylation
K309 Ubiquitination
T314 Phosphorylation
K369 Ubiquitination
K379 Ubiquitination
K384 Ubiquitination
K449 Ubiquitination
K533 Ubiquitination
S567 Phosphorylation
K616 Ubiquitination
T631 Phosphorylation
T665 Phosphorylation
S719 Phosphorylation
K777 Ubiquitination
Y861 Phosphorylation
T880 Phosphorylation
K898 Ubiquitination
K900 Ubiquitination
S909 Phosphorylation
S916 Phosphorylation
S918 Phosphorylation
S1131 Phosphorylation
K1133 Ubiquitination
T1162 Phosphorylation
S1166 Phosphorylation
S1171 Phosphorylation
T1172 Phosphorylation
K1187 Ubiquitination
Y1188 Phosphorylation
K1197 Ubiquitination
K1218 Acetylation
K1218 Ubiquitination
Y1232 Phosphorylation
K1256 Acetylation
K1256 Ubiquitination
K1257 Ubiquitination
S1261 Phosphorylation
T1262 Phosphorylation
K1267 Ubiquitination
S1288 Phosphorylation
K1293 Ubiquitination
S1297 Phosphorylation
S1299 Phosphorylation
K1395 Ubiquitination
K1400 Ubiquitination
K1406 Ubiquitination
S1415 Phosphorylation O15111 (CHUK)
S1418 Phosphorylation
K1465 Ubiquitination
K1471 Ubiquitination
K1500 Ubiquitination
T1502 Phosphorylation
K1511 Ubiquitination
K1566 Ubiquitination
K1635 Ubiquitination
K1655 Ubiquitination
K1662 Ubiquitination
K1745 Ubiquitination
Y1804 Phosphorylation
S1821 Phosphorylation
S1847 Phosphorylation
S1849 Phosphorylation
S1859 Phosphorylation
T1870 Phosphorylation
S1874 Phosphorylation
T1876 Phosphorylation
Y1880 Phosphorylation
S1893 Phosphorylation
T1948 Phosphorylation
K1993 Ubiquitination
K2045 Ubiquitination
K2066 Ubiquitination
S2069 Phosphorylation
S2155 Phosphorylation
S2159 Phosphorylation
T2164 Phosphorylation
K2166 Ubiquitination
T2173 Phosphorylation
K2218 Ubiquitination
K2301 Ubiquitination
K2370 Ubiquitination
T2380 Phosphorylation
T2434 Phosphorylation
T2436 Phosphorylation
S2442 Phosphorylation
T2444 Phosphorylation
T2446 Phosphorylation Q15418 (RPS6KA1) , P23443 (RPS6KB1) , Q13131 (PRKAA1) , P31749 (AKT1)
S2448 Phosphorylation P23443 (RPS6KB1) , P31749 (AKT1) , Q15418 (RPS6KA1)
Y2449 Phosphorylation
S2450 Phosphorylation
S2454 Phosphorylation P42345 (MTOR)
T2471 Phosphorylation
T2473 Phosphorylation P42345 (MTOR)
T2474 Phosphorylation P42345 (MTOR)
S2478 Phosphorylation P42345 (MTOR)
S2481 Phosphorylation P42345 (MTOR)
K2489 Ubiquitination
K2496 Ubiquitination

PTMs - P42345 As Enzyme

Substrate Site Source
O00141 (SGK1) S422 Uniprot
O00418 (EEF2K) S72 Uniprot
O00418 (EEF2K) S74 Uniprot
O75179 (ANKRD17) S2045 Uniprot
O75179 (ANKRD17) S2047 Uniprot
O75385 (ULK1) S638 Uniprot
O75385 (ULK1) S758 Uniprot
O95747 (OXSR1) S339 Uniprot
O95817 (BAG3) T285 Uniprot
O95817 (BAG3) S289 Uniprot
O96018 (APBA3) T5 Uniprot
O96018 (APBA3) S7 Uniprot
P01106 (MYC) S62 Uniprot
P03372 (ESR1) S104 Uniprot
P03372 (ESR1) S106 Uniprot
P04198 (MYCN) S62 Uniprot
P19484 (TFEB) S122 Uniprot
P19484 (TFEB) S142 Uniprot
P19484 (TFEB) S211 Uniprot
P23443-2 (RPS6KB1) T389 Uniprot
P23443 (RPS6KB1) T390 Uniprot
P23443 (RPS6KB1) S394 Uniprot
P23443 (RPS6KB1) T412 Uniprot
P23443 (RPS6KB1) S434 Uniprot
P23443 (RPS6KB1) S447 Uniprot
P26358 (DNMT1) S714 Uniprot
P31749 (AKT1) T450 Uniprot
P31749 (AKT1) S473 Uniprot
P35568 (IRS1) S307 Uniprot
P40763 (STAT3) S727 Uniprot
P42345 (MTOR) S2454 Uniprot
P42345 (MTOR) T2473 Uniprot
P42345 (MTOR) T2474 Uniprot
P42345 (MTOR) S2478 Uniprot
P42345 (MTOR) S2481 Uniprot
P51397 (DAP) S51 Uniprot
P55199 (ELL) S309 Uniprot
Q00613 (HSF1) S326 Uniprot
Q13322 (GRB10) T155 Uniprot
Q13322 (GRB10) S428 Uniprot
Q13322 (GRB10) S476 Uniprot
Q13541 (EIF4EBP1) T36 Uniprot
Q13541 (EIF4EBP1) T37 Uniprot
Q13541 (EIF4EBP1) T41 Uniprot
Q13541 (EIF4EBP1) S44 Uniprot
Q13541 (EIF4EBP1) T45 Uniprot
Q13541 (EIF4EBP1) T46 Uniprot
Q13541 (EIF4EBP1) S65 Uniprot
Q13541 (EIF4EBP1) T70 Uniprot
Q13541 (EIF4EBP1) S83 Uniprot
Q13541 (EIF4EBP1) S101 Uniprot
Q14693 (LPIN1) S106 Uniprot
Q14693 (LPIN1) S438 Uniprot
Q5T4S7 (UBR4) S2932 Uniprot
Q641Q2 (WASHC2A) S700 Uniprot
Q641Q2 (WASHC2A) S704 Uniprot
Q6PKG0 (LARP1) S766 Uniprot
Q6PKG0 (LARP1) S774 Uniprot
Q86TB9 (PATL1) S179 Uniprot
Q86TB9 (PATL1) S184 Uniprot
Q8IYB3 (SRRM1) T572 Uniprot
Q8IYB3 (SRRM1) T574 Uniprot
Q8N122 (RPTOR) S855 Uniprot
Q8N122 (RPTOR) S859 Uniprot
Q8N122 (RPTOR) S863 Uniprot
Q8TB45 (DEPTOR) S265 Uniprot
Q8TB45 (DEPTOR) S286 Uniprot
Q8TB45 (DEPTOR) S293 Uniprot
Q8TB45 (DEPTOR) T295 Uniprot
Q8TB45 (DEPTOR) S299 Uniprot
Q96B36 (AKT1S1) S183 Uniprot
Q96B36 (AKT1S1) S221 Uniprot
Q9BPZ7 (MAPKAP1) S260 Uniprot
Q9C0C7 (AMBRA1) S52 Uniprot
Q9H063 (MAF1) S60 Uniprot
Q9H063 (MAF1) S68 Uniprot
Q9H063 (MAF1) S75 Uniprot
Q9H1K1 (ISCU) S14 Uniprot
Q9H4A3 (WNK1) S2032 Uniprot
Q9H5H4 (ZNF768) S139 Uniprot
Q9H8M2 (BRD9) S588 Uniprot
Q9NQG5 (RPRD1B) S166 Uniprot
Q9P2Y5 (UVRAG) S550 Uniprot
Q9P2Y5 (UVRAG) S571 Uniprot
Q9UBS0-1 (RPS6KB2) T228 Uniprot
Q9UBS0-1 (RPS6KB2) S370 Uniprot
Q9UBS0 (RPS6KB2) T388 Uniprot
Q9UQ35 (SRRM2) S1318 Uniprot
Q9UQ35 (SRRM2) S1326 Uniprot
Q9UQ35 (SRRM2) S1329 Uniprot
Q9Y2J4 (AMOTL2) S759 Uniprot
Q9Y6W6 (DUSP10) S224 Uniprot
Q9Y6W6 (DUSP10) S230 Uniprot

Research Backgrounds

Function:

Serine/threonine protein kinase which is a central regulator of cellular metabolism, growth and survival in response to hormones, growth factors, nutrients, energy and stress signals. MTOR directly or indirectly regulates the phosphorylation of at least 800 proteins. Functions as part of 2 structurally and functionally distinct signaling complexes mTORC1 and mTORC2 (mTOR complex 1 and 2). Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. This includes phosphorylation of EIF4EBP1 and release of its inhibition toward the elongation initiation factor 4E (eiF4E) (By similarity). Moreover, phosphorylates and activates RPS6KB1 and RPS6KB2 that promote protein synthesis by modulating the activity of their downstream targets including ribosomal protein S6, eukaryotic translation initiation factor EIF4B, and the inhibitor of translation initiation PDCD4. Stimulates the pyrimidine biosynthesis pathway, both by acute regulation through RPS6KB1-mediated phosphorylation of the biosynthetic enzyme CAD, and delayed regulation, through transcriptional enhancement of the pentose phosphate pathway which produces 5-phosphoribosyl-1-pyrophosphate (PRPP), an allosteric activator of CAD at a later step in synthesis, this function is dependent on the mTORC1 complex. Regulates ribosome synthesis by activating RNA polymerase III-dependent transcription through phosphorylation and inhibition of MAF1 an RNA polymerase III-repressor. In parallel to protein synthesis, also regulates lipid synthesis through SREBF1/SREBP1 and LPIN1 (By similarity). To maintain energy homeostasis mTORC1 may also regulate mitochondrial biogenesis through regulation of PPARGC1A (By similarity). mTORC1 also negatively regulates autophagy through phosphorylation of ULK1 (By similarity). Under nutrient sufficiency, phosphorylates ULK1 at 'Ser-758', disrupting the interaction with AMPK and preventing activation of ULK1 (By similarity). Also prevents autophagy through phosphorylation of the autophagy inhibitor DAP. Also prevents autophagy by phosphorylating RUBCNL/Pacer under nutrient-rich conditions. mTORC1 exerts a feedback control on upstream growth factor signaling that includes phosphorylation and activation of GRB10 a INSR-dependent signaling suppressor. Among other potential targets mTORC1 may phosphorylate CLIP1 and regulate microtubules. As part of the mTORC2 complex MTOR may regulate other cellular processes including survival and organization of the cytoskeleton. Plays a critical role in the phosphorylation at 'Ser-473' of AKT1, a pro-survival effector of phosphoinositide 3-kinase, facilitating its activation by PDK1. mTORC2 may regulate the actin cytoskeleton, through phosphorylation of PRKCA, PXN and activation of the Rho-type guanine nucleotide exchange factors RHOA and RAC1A or RAC1B. mTORC2 also regulates the phosphorylation of SGK1 at 'Ser-422'. Regulates osteoclastogenesis by adjusting the expression of CEBPB isoforms (By similarity). Plays an important regulatory role in the circadian clock function; regulates period length and rhythm amplitude of the suprachiasmatic nucleus (SCN) and liver clocks (By similarity). Phosphorylates SQSTM1, promoting interaction between SQSTM1 and KEAP1 and subsequent inactivation of the BCR(KEAP1) complex (By similarity).

PTMs:

Autophosphorylates when part of mTORC1 or mTORC2. Phosphorylation at Ser-1261, Ser-2159 and Thr-2164 promotes autophosphorylation. Phosphorylation in the kinase domain modulates the interactions of MTOR with RPTOR and PRAS40 and leads to increased intrinsic mTORC1 kinase activity. Phosphorylation at Thr-2173 in the ATP-binding region by AKT1 strongly reduces kinase activity.

Subcellular Location:

Endoplasmic reticulum membrane>Peripheral membrane protein>Cytoplasmic side. Golgi apparatus membrane>Peripheral membrane protein>Cytoplasmic side. Mitochondrion outer membrane>Peripheral membrane protein>Cytoplasmic side. Lysosome. Cytoplasm. Nucleus>PML body. Microsome membrane. Lysosome membrane.
Note: Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB (PubMed:18497260, PubMed:20381137). Lysosome targeting also depends on interaction with MEAK7. Translocates to the lysosome membrane in the presence of TM4SF5 (PubMed:30956113).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in numerous tissues, with highest levels in testis.

Subunit Structure:

Part of the mammalian target of rapamycin complex 1 (mTORC1) which contains MTOR, MLST8, RPTOR, AKT1S1/PRAS40 and DEPTOR. The mTORC1 complex is a 1 Md obligate dimer of two stoichiometric heterotetramers with overall dimensions of 290 A x 210 A x 135 A. It has a rhomboid shape and a central cavity, the dimeric interfaces are formed by interlocking interactions between the two MTOR and the two RPTOR subunits. The MLST8 subunit forms distal foot-like protuberances, and contacts only one MTOR within the complex, while the small PRAS40 localizes to the midsection of the central core, in close proximity to RPTOR. Part of the mammalian target of rapamycin complex 2 (mTORC2) which contains MTOR, MLST8, PRR5, RICTOR, MAPKAP1 and DEPTOR. Interacts with PLPP7 and PML. Interacts with PRR5 and RICTOR; the interaction is direct within the mTORC2 complex. Interacts with WAC; WAC positively regulates MTOR activity by promoting the assembly of the TTT complex composed of TELO2, TTI1 and TTI2 and the RUVBL complex composed of RUVBL1 and RUVBL2 into the TTT-RUVBL complex which leads to the dimerization of the mTORC1 complex and its subsequent activation. Interacts with UBQLN1. Interacts with TTI1 and TELO2. Interacts with CLIP1; phosphorylates and regulates CLIP1. Interacts with NBN. Interacts with HTR6. Interacts with BRAT1. Interacts with MEAK7 (via C-terminal domain); the interaction increases upon nutrient stimulation. Interacts with TM4SF5; the interaction is positively regulated by arginine and is negatively regulated by leucine. Interacts with GPR137B.

Family&Domains:

The kinase domain (PI3K/PI4K) is intrinsically active but has a highly restricted catalytic center.

The FAT domain forms three discontinuous subdomains of alpha-helical TPR repeats plus a single subdomain of HEAT repeats. The four domains pack sequentially to form a C-shaped a-solenoid that clamps onto the kinase domain (PubMed:23636326).

Belongs to the PI3/PI4-kinase family.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - other.   (View pathway)

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Endocrine and metabolic diseases > Type II diabetes mellitus.

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Glioma.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway - multiple species.   (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

References

1). Aberrant translation regulated by METTL1/WDR4‐mediated tRNA N7‐methylguanosine modification drives head and neck squamous cell carcinoma progression. Cancer Communications, 2022 (PubMed: 35179319) [IF=16.2]

Application: WB    Species: Mice    Sample: METTL1‐KO cells

FIGURE 4 METTL1‐mediated m7G tRNA modification regulates the activity of the PI3K/AKT/mTOR signaling pathway. (A) Scatterplot of the TRs in METTL1‐WT and METTL1‐KO SCC15 cells. TRs were calculated by dividing the ribosome‐binding transcript signals by input RNA‐seq signals. (B) KEGG pathway analysis of the genes with decreased TRs upon METTL1 knockout. (C) The PI3K/AKT/mTOR signaling pathway was enriched in RNC‐seq datasets by GSEA (NES = 1.64, FDR = 0.165, P < 0.001). (D) Western blotting of PI3K/AKT/mTOR signaling pathway proteins and downstream proteins using the indicated antibodies. (E) qRT‐PCR analysis of PIK3CA with RNC and input samples in SCC9 and SCC15 cells. (F) The protein levels of PI3K, AKT, and p‐AKT in METTL1‐WT, METTL1‐KO, PI3K‐transfected METTL1‐KO cells (KO + PIK3CA) and 5 μg/mL SC79‐treated METTL1‐KO cells cultured with (KO + SC79). (G‐I) The proliferation (G), migration (H) and invasion abilities (I) were partially restored after transfecting METTL1‐KO cells with the PI3K plasmid or activating AKT. Data are presented as the mean ± SD and analyzed by Student's t‐test. *, P < 0.05, **, P < 0.01, ***, P < 0.001. Abbreviations: PI3K/AKT/mTOR: phosphatidylinositol‐3‐kinase/protein kinase B/mammalian target of rapamycin; METTL1: Methyltransferase‐like 1; WT: wild‐type; KO: knockout; TRs: translation ratios; KEGG: Koto Encyclopedia of Genes and Genomes; GSEA: gene set enrichment analysis; NES: normalized enrichment score; FDR: false discovery rate; qRT‐PCR: quantitative real‐time PCR; RNC: Ribosome nascent‐chain complex‐bound; MMP9: matrix metalloprotein 9; Bcl‐2: B‐cell lymphoma‐2; P‐S6K: phosphorylation of S6 kinase; BAX: Bcl‐2‐associated X protein; PIK3CA: phosphatidylinositol‐4,5‐bisphosphate 3‐kinase, catalytic subunit alpha; SD: standard deviation

2). Rationally designed rapamycin-encapsulated ZIF-8 nanosystem for overcoming chemotherapy resistance. BIOMATERIALS, 2020 (PubMed: 32841911) [IF=14.0]

Application: WB    Species: human    Sample: MCF-7/ADR cells

Figure 2.(b) ZIF-8 (10 µg mL−1, 24 h) did not inhibit p70S6K phosphorylation at Thr389 and mTOR phosphorylation at Ser2448 but induced the accumulation of LC3-II, suggesting ZIF-8 elicited mTOR-independent autophagy.

3). MiR-146b-5p enriched bioinspired exosomes derived from fucoidan-directed induction mesenchymal stem cells protect chondrocytes in osteoarthritis by targeting TRAF6. Journal of nanobiotechnology, 2023 (PubMed: 38105181) [IF=10.2]

Application: WB    Species: Rat    Sample: chondrocytes

Fig. 7 Enriched miR-146b-5p in F-MSCs-Exo inhibits PI3K/AKT/mTOR pathway by targeting TRAF6. (A, B) Western blot analysis was performed to detect the impact of F-MSCs-Exo on TRAF6 and the PI3K/AKT/mTOR pathway in rat chondrocytes. (C, D) The expression of TRAF6 was quantitatively analyzed using immunofluorescence staining and ImageJ software (scale bar = 10 μm). (E, F) Direct visualization of chondrocytes treated with nc-inhibitor and miR-146b-5p-inhibitor was performed using Alcian blue staining and safranin staining. (G, H) Western blot analysis was conducted to examine the expressions of TRAF6 and the PI3K/AKT/mTOR pathway in chondrocytes after treatment with nc-inhibitor and miR-146b-5p-inhibitor. (ns, no significant difference; *p 

4). NCAPD2 inhibits autophagy by regulating Ca2+/CAMKK2/AMPK/mTORC1 pathway and PARP-1/SIRT1 axis to promote colorectal cancer. CANCER LETTERS, 2021 (PubMed: 34229059) [IF=9.7]

Application: WB    Species: Human    Sample: CRC cells

Fig. 2. NCAPD2 inhibited cell autophagy and disrupted autophagic flux via Ca2+/CAMKK2/AMPK/mTORC1 pathway. (A) Western blot analyses for phosphorylated mTOR (p-mTOR, S2448), phosphorylated p70S6K (p-p70S6K, T389/412), phosphorylated 4E-BP1 (p-4E-BP1, T70) and phosphorylated AKT (p-AKT, S473) in CRCC cells with different treatments as indicated. (B) Western blot of indicated proteins in cells treated with mTORC1 inhibitor Rapamycin (3 nM, 24h). (C) Immunofluorescence staining of LC3II (red) and P62 (red) in CRC cells with different treatments as indicated. Merged images represented overlays of LC3II or P62 and nuclear staining by DAPI (blue). (D) Intracellular Ca2+ levels were analyzed by flow cytometry after staining with the fluorescent probe Fluo-3, AM in CRC cells. (E) Representative Western blot gel documents of phosphorylated CAMKK2(S511), phosphorylated AMPK(T172), phosphorylated mTOR(S2448), Beclin, ATG5, P62, LC3II in CRC cells with different treatments. (F) Western blots of indicated proteins in cells treated with an inhibitor of microsomal Ca2+-ATPase Thapsigargin (1 μM, 6h) and Ca2+ chelator BAPTA-AM (10 μM, 12h) respectively. Results are shown as mean ± s.d, *P < 0.05, **P < 0.01, ***P < 0.001, based on Student’s t-test. . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

5). Morphine-induced microglial immunosuppression via activation of insufficient mitophagy regulated by NLRX1. Journal of Neuroinflammation, 2022 (PubMed: 35414088) [IF=9.3]

6). Suppression of steroid 5α-reductase type I promotes cellular apoptosis and autophagy via PI3K/Akt/mTOR pathway in multiple myeloma. Cell Death & Disease, 2021 (PubMed: 33627630) [IF=9.0]

Application: WB    Species: human    Sample: ARP1 and H929 cells

Fig. 7 |Dutasteride possesses a therapeutic effect on MM.A MTT proliferation assay of ARP1 and H929 cells treated with different concentrations of dutasteride for 48 or 72 h. B Expression of apoptosis markers in dutasteride-treated ARP1 and H929 cells as determined by western blotting analysis. C Western blotting showed that Dutasteride treatment inhibited the activity of PI3K/Akt/mTOR signaling in ARP1 and H929 cells.

7). Polysaccharide from Strongylocentrotus nudus eggs regulates intestinal epithelial autophagy through CD36/PI3K-Akt pathway to ameliorate inflammatory bowel disease. International Journal of Biological Macromolecules, 2023 (PubMed: 37327932) [IF=8.2]

8). Cathepsin S activity controls chronic stress-induced muscle atrophy and dysfunction in mice. Cellular and Molecular Life Sciences, 2023 (PubMed: 37589754) [IF=8.0]

Application: WB    Species: Mouse    Sample:

Fig. 4 CTSS deficiency ameliorated stress-related anabolic and catabolic molecular alterations. a–e: Representative immunoblotting images and quantitative data for CTSS, IGF-1, IRS-2, p-PI3K, p-Akt, p-mTOR, p-FoxO1α, MuRF-1, MAFbx1, PGC-1α, PPAR-γ, C-caspase-3, and Bcl-2 in GAS muscles at Day 14 after stress (n = 3). Data are mean ± SEM, and p-values were determined by a one-way ANOVA followed by Bonferroni post hoc tests (b–e). CW: CTSS+/+ control mice, CK: CTSS−/− control mice, SW: 14-day-stressed CTSS+/+ mice, SK: 14-day-stressed CTSS−/− mice. *p 

9). Targeting mTOR/YY1 signaling pathway by quercetin through CYP7A1-mediated cholesterol-to-bile acids conversion alleviated type 2 diabetes mellitus induced hepatic lipid accumulation. Phytomedicine, 2023 (PubMed: 36889164) [IF=7.9]

10). Aiduqing formula suppresses breast cancer metastasis via inhibiting CXCL1-mediated autophagy. PHYTOMEDICINE, 2021 (PubMed: 34247114) [IF=7.9]

Application: WB    Species: human    Sample: MDA-MB-231 and BT-549 cells

Fig. 2. |ADQ formula suppresses autophagy and induces apoptosis of high-metastatic breast cancer cells. (A) The expression levels of autophagy-related proteins in MDA-MB-231 and BT-549 cells using western blotting analysis after treating with ADQ formula in a dose-dependent manner. Western-blotting results revealed that the expression levels of LC3-II, Beclin-1, and p-AMPK were downregulated while the expression levels of p62 and p-mTOR were upregulated in MDAMB-231 and BT-549 cells after ADQ treatment.

Application: WB    Species: Human    Sample: breast cancer cells

Fig. 2. ADQ formula suppresses autophagy and induces apoptosis of high-metastatic breast cancer cells. (A) The expression levels of autophagy-related proteins in MDA-MB-231 and BT-549 cells using western blotting analysis after treating with ADQ formula in a dose-dependent manner. Western-blotting results revealed that the expression levels of LC3-II, Beclin-1, and p-AMPK were downregulated while the expression levels of p62 and p-mTOR were upregulated in MDAMB-231 and BT-549 cells after ADQ treatment. (B) The abundance of autophagosomes and autolysosomes was measured with an LC3-GFP-mRFP reporter in MDAMB-231 and BT-549 cells after ADQ-formula treatment. The numbers of both yellow (autophagosomes) and red (autolysosomes) puncta were reduced in MDA-MB231 and BT-549 cells after ADQ-formula treatment. (C) The LC-3+ and early apoptosis subpopulations of MDA-MB-231 and BT-549 cells treat with ADQ formula for 24 h using flow cytometry. The result showed that ADQ formula significantly decreased LC-3+ subpopulations and promoted apoptosis in both MDA-MB-231 and BT549 cells. All values are presented as the mean ± SD, n = 3, ** p < 0.01, ## p < 0.01.

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