Akt1
Human Recombinant AKT1 - His,Strep Tagged
Reference ID:KB-3938
Western Blot
ELISA
FACS
Protein:Protein
Flow Cytometry
Gene of Interest
Gene Synonyms:AKT1;PKB;RAC
Protein Names:RAC-alpha serine/threonine-protein kinase (EC 2.7.11.1) (Protein kinase B) (PKB) (Protein kinase B alpha) (PKB alpha) (Proto-oncogene c-Akt) (RAC-PK-alpha)
Accession Data
Organism:Homo sapiens (Human)
Mass (kDa):556.86
Length (aa):480
Sequence:MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVDQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQTVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKVILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDRLCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLMLDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAKEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSMECVDSERRPHFPQFSYSASGTA
Proteomics (Proteome ID):UP000005640
Proteomics (Chromosome): Chromosome 14
Active Site:ACT_SITE 274 274 Proton acceptor. {ECO:0000255|PROSITE-ProRule:PRU00159, ECO:0000255|PROSITE-ProRule:PRU10027}.
Activity Regulation: Three specific sites, one in the kinase domain (Thr-308) and the two other ones in the C-terminal regulatory region (Ser-473 and Tyr-474), need to be phosphorylated for its full activation. Inhibited by pyrrolopyrimidine inhibitors like aniline triazole and spiroindoline. {ECO:0000269|PubMed:18456494, ECO:0000269|PubMed:20481595, ECO:0000269|PubMed:20810279, ECO:0000269|PubMed:21392984, ECO:0000269|PubMed:9512493, ECO:0000269|PubMed:9736715}.
Binding Site:BINDING 53 53 Inositol-(1,3,4,5)-tetrakisphosphate.; BINDING 86 86 Inositol-(1,3,4,5)-tetrakisphosphate.; BINDING 161 161 Inhibitor; via amide nitrogen.; BINDING 179 179 ATP. {ECO:0000255|PROSITE-ProRule:PRU00159}.; BINDING 230 230 Inhibitor; via amide nitrogen.; BINDING 234 234 Inhibitor.; BINDING 292 292 Inhibitor.
Catalytic Activity: Reaction=ATP + L-seryl-[protein] = ADP + H(+) + O-phospho-L-seryl-[protein]; Xref=Rhea:RHEA:17989, Rhea:RHEA-COMP:9863, Rhea:RHEA-COMP:11604, ChEBI:CHEBI:15378, ChEBI:CHEBI:29999, ChEBI:CHEBI:30616, ChEBI:CHEBI:83421, ChEBI:CHEBI:456216; EC=2.7.11.1; Evidence={ECO:0000269|PubMed:16139227, ECO:0000269|PubMed:1718748, ECO:0000269|PubMed:1851997}; Reaction=ATP + L-threonyl-[protein] = ADP + H(+) + O-phospho-L-threonyl-[protein]; Xref=Rhea:RHEA:46608, Rhea:RHEA-COMP:11060, Rhea:RHEA-COMP:11605, ChEBI:CHEBI:15378, ChEBI:CHEBI:30013, ChEBI:CHEBI:30616, ChEBI:CHEBI:61977, ChEBI:CHEBI:456216; EC=2.7.11.1; Evidence={ECO:0000269|PubMed:16139227, ECO:0000269|PubMed:1718748, ECO:0000269|PubMed:1851997};
Function [CC]:AKT1 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT regulates also the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity. Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven. AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319'. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates 'Ser-454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis. Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis. Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI(3)P-5 activity. The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development. Phosphorylates STK4/MST1 at 'Thr-120' and 'Thr-387' leading to inhibition of its: kinase activity, nuclear translocation, autophosphorylation and ability to phosphorylate FOXO3. Phosphorylates STK3/MST2 at 'Thr-117' and 'Thr-384' leading to inhibition of its: cleavage, kinase activity, autophosphorylation at Thr-180, binding to RASSF1 and nuclear translocation. Phosphorylates SRPK2 and enhances its kinase activity towards SRSF2 and ACIN1 and promotes its nuclear translocation. Phosphorylates RAF1 at 'Ser-259' and negatively regulates its activity. Phosphorylation of BAD stimulates its pro-apoptotic activity. Phosphorylates KAT6A at 'Thr-369' and this phosphorylation inhibits the interaction of KAT6A with PML and negatively regulates its acetylation activity towards p53/TP53.; AKT1-specific substrates have been recently identified, including palladin (PALLD), which phosphorylation modulates cytoskeletal organization and cell motility; prohibitin (PHB), playing an important role in cell metabolism and proliferation; and CDKN1A, for which phosphorylation at 'Thr-145' induces its release from CDK2 and cytoplasmic relocalization. These recent findings indicate that the AKT1 isoform has a more specific role in cell motility and proliferation. Phosphorylates CLK2 thereby controlling cell survival to ionizing radiation.
Kinetics:BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=52.8 uM for ATP (for purified and in vitro activated AKT1) {ECO:0000269|PubMed:16540465}; KM=0.5 uM for peptide substrate (for purified and in vitro activated AKT1) {ECO:0000269|PubMed:16540465}; KM=143.3 uM for ATP (for recombinant myristoylated AKT1 expressed and immunoprecipitated from Rat-1 cells) {ECO:0000269|PubMed:16540465}; KM=2.9 uM for peptide substrate (for recombinant myristoylated AKT1 expressed and immunoprecipitated from Rat-1 cells) {ECO:0000269|PubMed:16540465};
Nucleotide Binding:NP_BIND 156 164 ATP. {ECO:0000255|PROSITE-ProRule:PRU00159}.
Tissue Specificity:Expressed in prostate cancer and levels increase from the normal to the malignant state (at protein level). Expressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages. {ECO:0000269|PubMed:1718748, ECO:0000269|PubMed:17932490, ECO:0000269|PubMed:20333297}.
Chemical Profile | ChEBI:H(+) [CHEBI:15378]; ATP [CHEBI:30616]; O-phospho-L-threonine residue [CHEBI:61977]; O-phospho-L-serine residue [CHEBI:83421]; L-threonine residue [CHEBI:30013]; ADP [CHEBI:456216]; L-serine residue [CHEBI:29999]
ChEBI (Catalytic Activity):H(+) [CHEBI:15378]; ATP [CHEBI:30616]; O-phospho-L-threonine residue [CHEBI:61977]; O-phospho-L-serine residue [CHEBI:83421]; L-threonine residue [CHEBI:30013]; ADP [CHEBI:456216]; L-serine residue [CHEBI:29999]
Disease:Breast cancer (BC) [MIM:114480]: A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case. {ECO:0000269|PubMed:17611497}. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.; Colorectal cancer (CRC) [MIM:114500]: A complex disease characterized by malignant lesions arising from the inner wall of the large intestine (the colon) and the rectum. Genetic alterations are often associated with progression from premalignant lesion (adenoma) to invasive adenocarcinoma. Risk factors for cancer of the colon and rectum include colon polyps, long-standing ulcerative colitis, and genetic family history. Note=The gene represented in this entry may be involved in disease pathogenesis.; Note=Genetic variations in AKT1 may play a role in susceptibility to ovarian cancer.; Proteus syndrome (PROTEUSS) [MIM:176920]: A highly variable, severe disorder of asymmetric and disproportionate overgrowth of body parts, connective tissue nevi, epidermal nevi, dysregulated adipose tissue, and vascular malformations. Many features of Proteus syndrome overlap with other overgrowth syndromes. {ECO:0000269|PubMed:18954143, ECO:0000269|PubMed:21793738}. Note=The disease is caused by mutations affecting the gene represented in this entry.; Cowden syndrome 6 (CWS6) [MIM:615109]: A form of Cowden syndrome, a hamartomatous polyposis syndrome with age-related penetrance. Cowden syndrome is characterized by hamartomatous lesions affecting derivatives of ectodermal, mesodermal and endodermal layers, macrocephaly, facial trichilemmomas (benign tumors of the hair follicle infundibulum), acral keratoses, papillomatous papules, and elevated risk for development of several types of malignancy, particularly breast carcinoma in women and thyroid carcinoma in both men and women. Colon cancer and renal cell carcinoma have also been reported. Hamartomas can be found in virtually every organ, but most commonly in the skin, gastrointestinal tract, breast and thyroid. {ECO:0000269|PubMed:23246288}. Note=The disease is caused by mutations affecting the gene represented in this entry.
Mutagenesis:MUTAGEN 8 8 K->R: Substantial reduction of ubiquitination, phosphorylation at T-308 and S-473, AKT activation as well as IGF1-induced membrane recruitment. Decrease in ubiquitination and phosphorylation at T-308 as well as impaired association with the membrane; when associated with K-17. {ECO:0000269|PubMed:19713527}.; MUTAGEN 14 14 K->A: Impairs interaction with PtdIns(3,4,5)P3 and PtdIns(3,4)P2. {ECO:0000269|PubMed:12176338, ECO:0000269|PubMed:19713527, ECO:0000269|PubMed:21775285}.; MUTAGEN 14 14 K->Q: Substantial reduction of phosphorylation at T-308 and S-473, loss of AKT activation, and loss of binding to PIP3 as well as IGF1-induced membrane recruitment. {ECO:0000269|PubMed:12176338, ECO:0000269|PubMed:19713527, ECO:0000269|PubMed:21775285}.; MUTAGEN 14 14 K->R: Substantial reduction of ubiquitination, phosphorylation at T-308 and S-473, AKT activation, loss of binding to PIP3 as well as IGF1-induced membrane recruitment. {ECO:0000269|PubMed:12176338, ECO:0000269|PubMed:19713527, ECO:0000269|PubMed:21775285}.; MUTAGEN 17 17 E->K: No effect on membrane localization. Loss of membrane localization; when associated with Q-20. {ECO:0000269|PubMed:21775285}.; MUTAGEN 20 20 K->Q: Substantial reduction of phosphorylation at T-308 and S-473, reduced AKT activation, and reduced binding to PIP3 as well as IGF1-induced membrane recruitment. Loss of membrane localization; when associated with K-17. {ECO:0000269|PubMed:21775285}.; MUTAGEN 20 20 K->R: Slight increase of phosphorylation at T-308 and S-473. {ECO:0000269|PubMed:21775285}.; MUTAGEN 25 25 R->A,C: Impairs interaction with PtdIns(3,4,5)P3 and PtdIns(3,4)P2. {ECO:0000269|PubMed:12176338}.; MUTAGEN 86 86 R->A: Impairs interaction with PtdIns(3,4,5)P3 and PtdIns(3,4)P2. {ECO:0000269|PubMed:12176338}.; MUTAGEN 176 176 Y->F: Significant loss of interaction with TNK2. Loss of membrane localization. Significant reduction in phosphorylation on Ser-473. {ECO:0000269|PubMed:20333297}.; MUTAGEN 305 305 T->A: Reduces O-GlcNAc levels; Reduces O-GlcNAc levels even more; when associated with A-312. {ECO:0000269|PubMed:22629392}.; MUTAGEN 305 305 T->Y: Abolishes phosphorylation at Thr-308. {ECO:0000269|PubMed:22629392}.; MUTAGEN 308 308 T->D: 5-fold activation and 18-fold activation; when associated with D-473. {ECO:0000269|PubMed:12244301, ECO:0000269|PubMed:8978681}.; MUTAGEN 312 312 T->A: Reduces O-GlcNAc levels; Reduces O-GlcNAc levels even more; when associated with A-305. {ECO:0000269|PubMed:22629392}.; MUTAGEN 312 312 T->Y: Abolishes phosphorylation at Thr-308. {ECO:0000269|PubMed:22629392}.; MUTAGEN 473 473 S->D: 7-fold activation and 25-fold activation; when associated with D-308. {ECO:0000269|PubMed:12244301, ECO:0000269|PubMed:8978681}.; MUTAGEN 474 474 Y->F: 55% inhibition of activation. {ECO:0000269|PubMed:12149249}.
Reagent Data
Name:Akt1
Subcategory:Protein
Region:Met 1 - Ala 480
Molecular Weight:59.5
Source:HEK293
Species:Human
Tag:His Strep
Format:Lyophilized
Purification System:Chromatography
Formulation:Sterile-filtered colorless solution
Formulation Concentration:1 mg/ml
Buffer Volume:Standard
Buffer Solution:PBS
pH:7.4-7.5
Toxicity
Endotoxin Level:< 1%
Aggregate Tested By:SDS-PAGE
Endotoxin Screened:< 0.1 ng/ug
Purity:> 98%
Determined: SDS-PAGE
Stain:Blue
Chromotography:Anion-exchange
Validated: RP-HPLC
Sample Handling
Storage:-20°C
Stability:This bioreagent is stable at 4°C (short-term) and -70°C(long-term). After reconstitution, sample may be stored at 4°C for 2-7 days and below -18°C for future use.
Preparation:Reconstitute in sterile distilled H2O to no less than 100 ug/ml; dilute reconstituted stock further in other aqueous solutions if needed. Please review COA for lot-specific instructions. Final measurements should be determined by the end-user for optimal performance.