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Recombinant Human DNA PKcs protein (Tagged) is a Human Fragment protein, in the 3746 to 4128 aa range, expressed in Baculovirus infected Sf9, with >75% purity and suitable for SDS-PAGE.

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Images

SDS-PAGE - Recombinant Human DNA PKcs protein (Tagged) (AB204152), expandable thumbnail

Key facts

Purity
>75% Densitometry
Expression system
Baculovirus infected Sf9 cells
Tags
GST tag N-Terminus
Applications
SDS-PAGE
Biologically active
No

Amino acid sequence

R E H P F L V K G G E D L R Q D Q R V E Q L F Q V M N G I L A Q D S A C S Q R A L Q L R T Y S V V P M T S R L G L I E W L E N T V T L K D L L L N T M S Q E E K A A Y L S D P R A P P C E Y K D W L T K M S G K H D V G A Y M L M Y K G A N R T E T V T S F R K R E S K V P A D L L K R A F V R M S T S P E A F L A L R S H F A S S H A L I C I S H W I L G I G D R H L N N F M V A M E T G G V I G I D F G H A F G S A T Q F L P V P E L M P F R L T R Q F I N L M L P M K E T G L M Y S I M V H A L R A F R S D P G L L T N T M D V F V K E P S F D W K N F E Q K M L K K G G S W I Q E I N V A E K N W Y P R Q K I C Y A K R K L A G A N P A V I T C D E L L L G H E K A P A F R D Y V A V A R G S K D H N I R A Q E P E S G L S E E T Q V K C L M D Q A T D P N I L G R T W E G W E P W M

Reactivity data

Application
SDS-PAGE
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:33854234). Involved in DNA non-homologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:33854234, PubMed:34352203). Must be bound to DNA to express its catalytic properties (PubMed:11955432). Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C) (PubMed:11955432). Recruited by XRCC5 and XRCC6 to DNA ends and is required to (1) protect and align broken ends of DNA, thereby preventing their degradation, (2) and sequester the DSB for repair by NHEJ (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326, PubMed:33854234). Act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion (By similarity). Also involved in modulation of transcription (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). As part of the DNA-PK complex, involved in the early steps of ribosome assembly by promoting the processing of precursor rRNA into mature 18S rRNA in the small-subunit processome (PubMed:32103174). Binding to U3 small nucleolar RNA, recruits PRKDC and XRCC5/Ku86 to the small-subunit processome (PubMed:32103174). Recognizes the substrate consensus sequence [ST]-Q (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). Phosphorylates 'Ser-139' of histone variant H2AX, thereby regulating DNA damage response mechanism (PubMed:14627815, PubMed:16046194). Phosphorylates ASF1A, DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, FH, SRF, NHEJ1/XLF, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2 (PubMed:10026262, PubMed:10467406, PubMed:11889123, PubMed:12509254, PubMed:14599745, PubMed:14612514, PubMed:14704337, PubMed:15177042, PubMed:1597196, PubMed:16397295, PubMed:18644470, PubMed:2247066, PubMed:2507541, PubMed:26237645, PubMed:26666690, PubMed:28712728, PubMed:29478807, PubMed:30247612, PubMed:8407951, PubMed:8464713, PubMed:9139719, PubMed:9362500). Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA (PubMed:9679063). Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D (PubMed:9363941). Contributes to the determination of the circadian period length by antagonizing phosphorylation of CRY1 'Ser-588' and increasing CRY1 protein stability, most likely through an indirect mechanism (By similarity). Plays a role in the regulation of DNA virus-mediated innate immune response by assembling into the HDP-RNP complex, a complex that serves as a platform for IRF3 phosphorylation and subsequent innate immune response activation through the cGAS-STING pathway (PubMed:28712728). Also regulates the cGAS-STING pathway by catalyzing phosphorylation of CGAS, thereby impairing CGAS oligomerization and activation (PubMed:33273464). Also regulates the cGAS-STING pathway by mediating phosphorylation of PARP1 (PubMed:35460603).

Alternative names

Recommended products

Recombinant Human DNA PKcs protein (Tagged) is a Human Fragment protein, in the 3746 to 4128 aa range, expressed in Baculovirus infected Sf9, with >75% purity and suitable for SDS-PAGE.

Key facts

Purity
>75% Densitometry
Expression system
Baculovirus infected Sf9 cells
Applications
SDS-PAGE
Accession
P78527-1
Animal free
No
Species
Human
Concentration
Loading...
Storage buffer

pH: 7.5
Constituents: 25% Glycerol (glycerin, glycerine), 0.88% Sodium chloride, 0.79% Tris HCl, 0.31% Glutathione, 0.004% (R*,R*)-1,4-Dimercaptobutan-2,3-diol, 0.003% EDTA, 0.002% PMSF

Sequence info

Amino acid sequence

R E H P F L V K G G E D L R Q D Q R V E Q L F Q V M N G I L A Q D S A C S Q R A L Q L R T Y S V V P M T S R L G L I E W L E N T V T L K D L L L N T M S Q E E K A A Y L S D P R A P P C E Y K D W L T K M S G K H D V G A Y M L M Y K G A N R T E T V T S F R K R E S K V P A D L L K R A F V R M S T S P E A F L A L R S H F A S S H A L I C I S H W I L G I G D R H L N N F M V A M E T G G V I G I D F G H A F G S A T Q F L P V P E L M P F R L T R Q F I N L M L P M K E T G L M Y S I M V H A L R A F R S D P G L L T N T M D V F V K E P S F D W K N F E Q K M L K K G G S W I Q E I N V A E K N W Y P R Q K I C Y A K R K L A G A N P A V I T C D E L L L G H E K A P A F R D Y V A V A R G S K D H N I R A Q E P E S G L S E E T Q V K C L M D Q A T D P N I L G R T W E G W E P W M
Accession
P78527
Protein length
Fragment
Predicted molecular weight
68 kDa
Amino acids
3746 to 4128
Nature
Recombinant
Tags
GST tag N-Terminus

Specifications

Form
Liquid
Additional notes

Affinity purified.

General info

Function

Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:33854234). Involved in DNA non-homologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:33854234, PubMed:34352203). Must be bound to DNA to express its catalytic properties (PubMed:11955432). Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C) (PubMed:11955432). Recruited by XRCC5 and XRCC6 to DNA ends and is required to (1) protect and align broken ends of DNA, thereby preventing their degradation, (2) and sequester the DSB for repair by NHEJ (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326, PubMed:33854234). Act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion (By similarity). Also involved in modulation of transcription (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). As part of the DNA-PK complex, involved in the early steps of ribosome assembly by promoting the processing of precursor rRNA into mature 18S rRNA in the small-subunit processome (PubMed:32103174). Binding to U3 small nucleolar RNA, recruits PRKDC and XRCC5/Ku86 to the small-subunit processome (PubMed:32103174). Recognizes the substrate consensus sequence [ST]-Q (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). Phosphorylates 'Ser-139' of histone variant H2AX, thereby regulating DNA damage response mechanism (PubMed:14627815, PubMed:16046194). Phosphorylates ASF1A, DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, FH, SRF, NHEJ1/XLF, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2 (PubMed:10026262, PubMed:10467406, PubMed:11889123, PubMed:12509254, PubMed:14599745, PubMed:14612514, PubMed:14704337, PubMed:15177042, PubMed:1597196, PubMed:16397295, PubMed:18644470, PubMed:2247066, PubMed:2507541, PubMed:26237645, PubMed:26666690, PubMed:28712728, PubMed:29478807, PubMed:30247612, PubMed:8407951, PubMed:8464713, PubMed:9139719, PubMed:9362500). Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA (PubMed:9679063). Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D (PubMed:9363941). Contributes to the determination of the circadian period length by antagonizing phosphorylation of CRY1 'Ser-588' and increasing CRY1 protein stability, most likely through an indirect mechanism (By similarity). Plays a role in the regulation of DNA virus-mediated innate immune response by assembling into the HDP-RNP complex, a complex that serves as a platform for IRF3 phosphorylation and subsequent innate immune response activation through the cGAS-STING pathway (PubMed:28712728). Also regulates the cGAS-STING pathway by catalyzing phosphorylation of CGAS, thereby impairing CGAS oligomerization and activation (PubMed:33273464). Also regulates the cGAS-STING pathway by mediating phosphorylation of PARP1 (PubMed:35460603).

Sequence similarities

Belongs to the PI3/PI4-kinase family.

Post-translational modifications

Autophosphorylated at two clusters, the T2609 cluster and the S2056 cluster (PubMed:33854234). Autophosphorylated on Ser-2056, Thr-2609, Thr-2638 and Thr-2647 (PubMed:12186630, PubMed:12231622, PubMed:14734805, PubMed:33854234). Ser-2056 and Thr-2609 are DNA damage-inducible phosphorylation sites (inducible with ionizing radiation, IR) dephosphorylated by PPP5C (PubMed:12186630, PubMed:12231622, PubMed:14734805). Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, requisite for efficient end processing and DNA repair (PubMed:12186630, PubMed:12231622, PubMed:14734805). Autophosphorylation in trans within DNA-PK complexes loaded on DNA ends leads to the dissociation of PRKDC from DNA and the transition into the short-range NHEJ complex (PubMed:33854234). Autophosphorylation of the T2609 cluster is required for hematopoietic development and protein synthesis in erythrocytes precursors (By similarity).

Subcellular localisation
Nucleus, Nucleolus

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-80°C
Appropriate long-term storage conditions
-80°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

DNA-dependent protein kinase catalytic subunit often abbreviated as DNA-PKcs or PKcs is an important component in the cellular machinery responsible for DNA repair. This protein weighs approximately 470 kDa and is predominantly expressed in various tissue types with higher levels found in lymphoid tissues. The DNA-PKcs functions as a serine/threonine protein kinase and becomes active upon binding to DNA contributing to its role in maintaining genome integrity.

Biological function summary

DNA-PKcs plays an essential role in the non-homologous end joining (NHEJ) repair pathway which repairs double-strand breaks in DNA. DNA-PKcs forms a complex with the Ku70/80 heterodimer serving as a critical component of the DNA repair mechanism. This complex recognizes DNA ends facilitates their synapsis and activates other enzymes involved in ligating the broken DNA strands therefore promoting cellular survival following DNA damage.

Pathways

DNA-PKcs deeply integrates into the DNA damage response pathway. It collaborates closely with proteins such as ATM (ataxia-telangiectasia mutated) to sense DNA damage and initiate repair. DNA-PKcs is involved in V(D)J recombination vital for the generation of diverse antibodies in immune cells. Its activity is essential for coordinating with other proteins including XRCC4 and Ligase IV to ensure efficient DNA repair processes are executed.

Associated diseases and disorders

Defects in DNA-PKcs are linked to severe combined immunodeficiency (SCID) due to its central role in V(D)J recombination. Dysfunction in its pathway also relates to cancer where impaired DNA repair mechanisms substantially increase genomic instability leading to tumorigenesis. DNA-PKcs interacts with other proteins like BRCA1 and p53 in tumors where its overexpression or mutations can contribute to resistance against radiation and chemotherapeutic agents. These interactions highlight the importance of DNA-PK inhibitors as potential therapeutic strategies in oncology.

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1 product image

  • SDS-PAGE - Recombinant Human DNA PKcs protein (Tagged) (ab204152), expandable thumbnail

    SDS-PAGE - Recombinant Human DNA PKcs protein (Tagged) (ab204152)

    SDS-PAGE analysis of ab204512 which was determined to be >75% pure by densitometry.

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