Phospho-DEPDC5 (Ser1530) Antibody - #AF8617

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製品: Phospho-DEPDC5 (Ser1530) Antibody
カタログ: AF8617
タンパク質の説明: Rabbit polyclonal antibody to Phospho-DEPDC5 (Ser1530)
アプリケーション: WB
反応性: Human
予測: Zebrafish, Bovine, Horse, Rabbit, Dog, Chicken
分子量: 180 kDa; 181kD(Calculated).
ユニプロット: O75140
RRID: AB_2844377

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製品説明

ソース:
Rabbit
アプリケーション:
WB 1:1000-3000
*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.

反応性:
Human
予測:
Zebrafish(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%)
クローナリティ:
Polyclonal
特異性:
Phospho-DEPDC5 (Ser1530) Antibody detects endogenous levels of DEPDC5 only when phosphorylated at S1530.
RRID:
AB_2844377
引用形式: Affinity Biosciences Cat# AF8617, RRID:AB_2844377.
コンジュゲート:
Unconjugated.
精製:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
保存:
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.
別名:

折りたたみ／展開

DEP domain containing 5; DEP domain-containing protein 5; DEP.5; DEPD5_HUMAN; DEPDC5; FFEVF; KIAA0645;

免疫原

免疫原:
Uniprot:

O75140(DEPD5_HUMAN) >>Visit HPA database.

遺伝子(ID):
DEPDC5(9681)
発現特異性:
O75140 DEPD5_HUMAN:

Expressed in developing and adult brain.

タンパク質配列:
MRTTKVYKLVIHKKGFGGSDDELVVNPKVFPHIKLGDIVEIAHPNDEYSPLLLQVKSLKEDLQKETISVDQTVTQVFRLRPYQDVYVNVVDPKDVTLDLVELTFKDQYIGRGDMWRLKKSLVSTCAYITQKVEFAGIRAQAGELWVKNEKVMCGYISEDTRVVFRSTSAMVYIFIQMSCEMWDFDIYGDLYFEKAVNGFLADLFTKWKEKNCSHEVTVVLFSRTFYDAKSVDEFPEINRASIRQDHKGRFYEDFYKVVVQNERREEWTSLLVTIKKLFIQYPVLVRLEQAEGFPQGDNSTSAQGNYLEAINLSFNVFDKHYINRNFDRTGQMSVVITPGVGVFEVDRLLMILTKQRMIDNGIGVDLVCMGEQPLHAVPLFKLHNRSAPRDSRLGDDYNIPHWINHSFYTSKSQLFCNSFTPRIKLAGKKPASEKAKNGRDTSLGSPKESENALPIQVDYDAYDAQVFRLPGPSRAQCLTTCRSVRERESHSRKSASSCDVSSSPSLPSRTLPTEEVRSQASDDSSLGKSANILMIPHPHLHQYEVSSSLGYTSTRDVLENMMEPPQRDSSAPGRFHVGSAESMLHVRPGGYTPQRALINPFAPSRMPMKLTSNRRRWMHTFPVGPSGEAIQIHHQTRQNMAELQGSGQRDPTHSSAELLELAYHEAAGRHSNSRQPGDGMSFLNFSGTEELSVGLLSNSGAGMNPRTQNKDSLEDSVSTSPDPILTLSAPPVVPGFCCTVGVDWKSLTTPACLPLTTDYFPDRQGLQNDYTEGCYDLLPEADIDRRDEDGVQMTAQQVFEEFICQRLMQGYQIIVQPKTQKPNPAVPPPLSSSPLYSRGLVSRNRPEEEDQYWLSMGRTFHKVTLKDKMITVTRYLPKYPYESAQIHYTYSLCPSHSDSEFVSCWVEFSHERLEEYKWNYLDQYICSAGSEDFSLIESLKFWRTRFLLLPACVTATKRITEGEAHCDIYGDRPRADEDEWQLLDGFVRFVEGLNRIRRRHRSDRMMRKGTAMKGLQMTGPISTHSLESTAPPVGKKGTSALSALLEMEASQKCLGEQQAAVHGGKSSAQSAESSSVAMTPTYMDSPRKDGAFFMEFVRSPRTASSAFYPQVSVDQTATPMLDGTSLGICTGQSMDRGNSQTFGNSQNIGEQGYSSTNSSDSSSQQLVASSLTSSSTLTEILEAMKHPSTGVQLLSEQKGLSPYCFISAEVVHWLVNHVEGIQTQAMAIDIMQKMLEEQLITHASGEAWRTFIYGFYFYKIVTDKEPDRVAMQQPATTWHTAGVDDFASFQRKWFEVAFVAEELVHSEIPAFLLPWLPSRPASYASRHSSFSRSFGGRSQAAALLAATVPEQRTVTLDVDVNNRTDRLEWCSCYYHGNFSLNAAFEIKLHWMAVTAAVLFEMVQGWHRKATSCGFLLVPVLEGPFALPSYLYGDPLRAQLFIPLNISCLLKEGSEHLFDSFEPETYWDRMHLFQEAIAHRFGFVQDKYSASAFNFPAENKPQYIHVTGTVFLQLPYSKRKFSGQQRRRRNSTSSTNQNMFCEERVGYNWAYNTMLTKTWRSSATGDEKFADRLLKDFTDFCINRDNRLVTFWTSCLEKMHASAP

種類予測

種類予測:

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

PTMs - O75140 基板として

Site PTM Type Enzyme
T66 Phosphorylation
Y155 Phosphorylation
T273 Phosphorylation
K411 Methylation
S432 Phosphorylation
S445 Phosphorylation
S489 Phosphorylation
S491 Phosphorylation
S494 Phosphorylation
S496 Phosphorylation
S497 Phosphorylation
S503 Phosphorylation
S505 Phosphorylation
S508 Phosphorylation
S521 Phosphorylation
S524 Phosphorylation
S525 Phosphorylation
S548 Phosphorylation
S579 Phosphorylation
T749 Phosphorylation
Y759 Phosphorylation
S831 Phosphorylation
S833 Phosphorylation
Y920 Phosphorylation
Y924 Phosphorylation
S1028 Phosphorylation
S1530 Phosphorylation
T1531 Phosphorylation
Y1546 Phosphorylation
Y1550 Phosphorylation
K1574 Ubiquitination

研究背景

機能:

As a component of the GATOR1 complex functions as an inhibitor of the amino acid-sensing branch of the TORC1 pathway. The GATOR1 complex strongly increases GTP hydrolysis by RRAGA and RRAGB within RRAGC-containing heterodimers, thereby deactivating RRAGs, releasing mTORC1 from lysosomal surface and inhibiting mTORC1 signaling. The GATOR1 complex is negatively regulated by GATOR2 the other GATOR subcomplex in this amino acid-sensing branch of the TORC1 pathway.

PTMs:

Ubiquitinated. Amino acid-induced 'Lys-48'-linked polyubiquitination of DEPDC5 by the BCR(KLHL22) ubiquitin ligase complex leads to DEPDC5 proteasomal degradation and inhibition of the GATOR1 complex. Ubiquitination may occur at multiple lysines.

細胞の位置付け:

Cytoplasm>Cytosol. Cytoplasm>Perinuclear region. Lysosome membrane.
Note: Localization to lysosomes is amino acid-independent.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Expressed in developing and adult brain.

サブユニット構造:

Within the GATOR complex, component of the GATOR1 subcomplex, made of DEPDC5, NPRL2 and NPRL3. GATOR1 mediates the strong interaction of the GATOR complex with RRAGA/RRAGC and RRAGB/RRAGC heterodimers. Interacts (via DEP domain) with KLHL22; the interaction depends on amino acid availability.

タンパク質ファミリー:

The DEP domain mediates the interaction with KLHL22.

Belongs to the IML1 family.

研究領域

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

参考文献

1). Phosphorylation of DEPDC5, a component of the GATOR1 complex, releases inhibition of mTORC1 and promotes tumor growth. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2019 (PubMed: 31548394) [IF=11.1]

Application: WB    Species: Human    Sample: 293T cells

Fig. 2. Pim and AKT kinases phosphorylate DEPDC5. (A) Pim1 and Flag-tagged DEPDC5 (Flag DEPDC5) plasmids were cotransfected into 293T cells. Flag immunoprecipitated (IP) samples and whole cell lysates (WCL) were analyzed by WB using the indicated antibodies. (B and C) Plasmids expressing DEPDC5 WT, S1002A (SA1), and S1530A (SA2) were transfected into 293T cells which were treated with and without 3 µM Pim447 for 16 h and then cells were lysed. WCL and Flag-IP samples were analyzed by WB using the indicated antibodies. (D and E) Plasmids expressing Flag DEPDC5, Pim1, or myr-AKT were cotransfected into 293T cells and then treated with 3 µM Pim447 or the AKT inhibitor AZD5363, 3 µM for 18 h. The Flag-IP samples and WCL were analyzed by WB. (F) BT549 cells stably expressing EGFP as a control or DEPDC5 were incubated with DMSO (C), 3 µM Pim447 (P), 3 µM AZD5363 (A), and the combination (AP) for 6 h and cell lysates were analyzed by WB using the indicated antibodies. (G) BT549 cells stably expressing DEPDC5 were starved for 24 h and pretreated with DMSO (C), 3 µM Pim447 (P), 5 µM GSK690693 (A), and the combination (AP) for 1.5 h before stimulation with insulin (0.5 µg/mL) for 30 min, and cell lysates were analyzed by WB using the indicated antibodies. (H) BT549 cells stably expressing DEPDC5 were transfected with siRNA targeting Pim1 or all 3 Pim kinases, and after 48 h, cell lysates were analyzed by WB. See SI Appendix, SI Materials and Methods for WB quantification. Numerical values shown are calculated relative to the DMSO (C) treatment for F or no insulin treatment for G.

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