Active Recombinant Full Length Human PIK3CA(E545K) and PIK3R1 Mutation Co-expressed Protein, N-His-tagged

• Cat.No.: PIK3CA-PIK3R1-129HFL
• Product Overview: Complex of N-terminal 6His-tagged, recombinant, full-length human p110α containing the mutation E545K and untagged, recombinant, full-length human p85α. Co-expressed by baculovirus in Sf21 insect cells and purified using Ni2+/NTA-agarose. The E545K substitution is a somatic mutation in p110α that has been associated with tumours of the colon and brain. Combined in vitro and in vivo studies have shown that this mutation confers higher lipid kinase activity than wild type, and is able to induce oncogenic transformation.

Specification

• Description: Complex of N-terminal 6His-tagged recombinant, human full length p110α containing the mutation E545K and untagged, recombinant, full length human p85α. The E545K substitution is a somatic mutation in p110α that has been associated with tumors of the colon and brain. Combined in vitro and in vivo studies have shown that this mutation confers higher lipid kinase activity than wild type, and is able to induce oncogenic transformation.
• Source: Sf21
• Species: Human
• Tag: His
• Protein length: Full Length
• Form: 0.789mg/ml of enzyme in 50mM Tris/HCl pH8.0, 300mM NaCl, 0.1mM EGTA, 0.03% Brij-35, 270mM sucrose, 0.2mM PMSF, 1mM benzamidine, 0.1% 2- mercaptoethanol. Frozen solution.
• Bio-activity: 355U/mg, where one unit of PI 3-Kinase alpha (p110α(E545K)/p85α (h)) activity is defined as 1nmol phosphatidylinositol 3,4,5-trisphosphate (PIP3) formed per minute at room temperature with a final ATP concentration of 100μM.
• Molecular Mass: p110α(E545K) MW = 125.3kDa, p85α MW = 83.7kDa
• AA Sequence: p110α(E545K) amino acid sequence: 1 MHHHHHHMPP RPSSGELWGI HLMPPRILVE CLLPNGMIVT LECLREATLI TIKHELFKEA 61 RKYPLHQLLQ DESSYIFVSV TQEAEREEFF DETRRLCDLR LFQPFLKVIE PVGNREEKIL 121 NREIGFAIGM PVCEFDMVKD PEVQDFRRNI LNVCKEAVDL RDLNSPHSRA MYVYPPNVES 181 SPELPKHIYN KLDKGQIIVV IWVIVSPNND KQKYTLKINH DCVPEQVIAE AIRKKTRSML 241 LSSEQLKLCV LEYQGKYILK VCGCDEYFLE KYPLSQYKYI RSCIMLGRMP NLMLMAKESL 301 YSQLPMDCFT MPSYSRRIST ATPYMNGETS TKSLWVINSA LRIKILCATY VNVNIRDIDK 361 IYVRTGIYHG GEPLCDNVNT QRVPCSNPRW NEWLNYDIYI PDLPRAARLC LSICSVKGRK 421 GAKEEHCPLA WGNINLFDYT DTLVSGKMAL NLWPVPHGLE DLLNPIGVTG SNPNKETPCL 481 ELEFDWFSSV VKFPDMSVIE EHANWSVSRE AGFSYSHAGL SNRLARDNEL RENDKEQLKA 541 ISTRDPLSEI TKQEKDFLWS HRHYCVTIPE ILPKLLLSVK WNSRDEVAQM YCLVKDWPPI 601 KPEQAMELLD CNYPDPMVRG FAVRCLEKYL TDDKLSQYLI QLVQVLKYEQ YLDNLLVRFL 661 LKKALTNQRI GHFFFWHLKS EMHNKTVSQR FGLLLESYCR ACGMYLKHLN RQVEAMEKLI 721 NLTDILKQEK KDETQKVQMK FLVEQMRRPD FMDALQGFLS PLNPAHQLGN LRLEECRIMS 781 SAKRPLWLNW ENPDIMSELL FQNNEIIFKN GDDLRQDMLT LQIIRIMENI WQNQGLDLRM 841 LPYGCLSIGD CVGLIEVVRN SHTIMQIQCK GGLKGALQFN SHTLHQWLKD KNKGEIYDAA 901 IDLFTRSCAG YCVATFILGI GDRHNSNIMV KDDGQLFHID FGHFLDHKKK KFGYKRERVP 961 FVLTQDFLIV ISKGAQECTK TREFERFQEM CYKAYLAIRQ HANLFINLFS MMLGSGMPEL 1021 QSFDDIAYIR KTLALDKTEQ EALEYFMKQM NDAHHGGWTT KMDWIFHTIK QHALN
  p85α amino acid sequence:
  1 MSAEGYQYRA LYDYKKEREE DIDLHLGDIL TVNKGSLVAL GFSDGQEARP EEIGWLNGYN
  61 ETTGERGDFP GTYVEYIGRK KISPPTPKPR PPRPLPVAPG SSKTEADVEQ QALTLPDLAE
  121 QFAPPDIAPP LLIKLVEAIE KKGLECSTLY RTQSSSNLAE LRQLLDCDTP SVDLEMIDVH
  181 VLADAFKRYL LDLPNPVIPA AVYSEMISLA PEVQSSEEYI QLLKKLIRSP SIPHQYWLTL
  241 QYLLKHFFKL SQTSSKNLLN ARVLSEIFSP MLFRFSAASS DNTENLIKVI EILISTEWNE
  301 RQPAPALPPK PPKPTTVANN GMNNNMSLQD AEWYWGDISR EEVNEKLRDT ADGTFLVRDA
  361 STKMHGDYTL TLRKGGNNKL IKIFHRDGKY GFSDPLTFSS VVELINHYRN ESLAQYNPKL
  421 DVKLLYPVSK YQQDQVVKED NIEAVGKKLH EYNTQFQEKS REYDRLYEEY TRTSQEIQMK
  481 RTAIEAFNET IKIFEEQCQT QERYSKEYIE KFKREGNEKE IQRIMHNYDK LKSRISEIID
  541 SRRRLEEDLK KQAAEYREID KRMNSIKPDL IQLRKTRDQY LMWLTQKGVR QKKLNEWLGN
  601 ENTEDQYSLV EDDEDLPHHD EKTWNVGSSN RNKAENLLRG KRDGTFLVRE SSKQGCYACS
  661 VVVDGEVKHC VINKTATGYG FAEPYNLYSS LKELVLHYQH TSLVQHNDSL NVTLAYPVYA
  721 QQRR
• Purity: Purity (p110α / p85α combined) 85.8% by SDS-PAGE and Coomassie blue staining.
• Applications: Drug Discovery & Development
• Storage: 6 months at -70 centigrade.
• Handling Advice: Rapidly thaw the vial under cold water and immediately place on ice. Aliquot unused material into prechilled micro-centrifuge tubes and immediately snap-freeze the vials in liquid nitrogen prior to re-storage at -70 centigrade.
• Use/Stability: On receipt of material store at -70 centigrade. Unopened reagent is stable for a minimum of 1 year from date of shipment when stored at recommended storage temperature. Avoid repeat freeze/thaw cycles. For maximum recovery of product, centrifuge original vial prior to removing the cap.
• References: Kang S. et al., PNAS, (2005);102: 802-807 and Zhao J.J. et al., PNAS, (2005);102:18443-18448.

Gene Information

• Gene Name: PIK3CA phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha [ Homo sapiens (human) ] and PIK3R1 phosphoinositide-3-kinase regulatory subunit 1 [ Homo sapiens (human) ]
• Official Symbol: PIK3CA and PIK3R1
• Synonyms: PIK3CA and PIK3R1; phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha and phosphoinositide-3-kinase regulatory subunit 1; MCM; CCM4; CWS5; MCAP; PI3K; CLAPO; CLOVE; MCMTC; PI3K-alpha; p110-alpha and p85; AGM7; GRB1; IMD36; p85alpha; p85-ALPHA
• Gene ID: 5290 and 5295
• mRNA Refseq: NM_006218.4 and NM_181504.4
• Protein Refseq: NP_006209.2 and NP_852556.2
• MIM: 171834 and 171833
• UniProt ID: P42336 and A0A2X0SFG1