Phospho-MKK3/MKK6 (Ser189/Thr193)) Antibody - #AF3805

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Product: Phospho-MKK3/MKK6 (Ser189/Thr193)) Antibody
Catalog: AF3805
Description: Rabbit polyclonal antibody to Phospho-MKK3/MKK6 (Ser189/Thr193))
Application: WB IHC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 39kD,37kD(Calculated).
Uniprot: P46734 | P52564
RRID: AB_2847119

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
IHC 1:50-1:200, WB 1:500-1:2000
*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
Clonality:
Polyclonal
Specificity:
Phospho-MKK3/MKK6 (Ser189/Thr193)) Antibody detects endogenous levels of MKK3/MKK6 only when phosphorylated at Ser189/Thr193.
RRID:
AB_2847119
Cite Format: Affinity Biosciences Cat# AF3805, RRID:AB_2847119.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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

AW212142; dual specificity mitogen activated protein kinase kinase 3; Dual specificity mitogen-activated protein kinase kinase 3; MAP kinase kinase 3; map2k3; MAPK ERK kinase 3; MAPK/ERK kinase 3; MAPKK 3; MAPKK3; MEK 3; MEK3; Mitogen activated protein kinase kinase 3; MKK 3; MKK3; mMKK3b; MP2K3_HUMAN; PRKMK 3; PRKMK3; protein kinase, mitogen-activated, kinase 3; SAPK kinase 2; SAPKK 2; SAPKK2; Stress activated protein kinase kinase 2; Dual specificity mitogen activated protein kinase kinase 6; Dual specificity mitogen-activated protein kinase kinase 6; MAP kinase kinase 6; MAP2K6; MAPK/ERK kinase 6; MAPKK 6; MAPKK6; MEK 6; MEK6; Mitogen Activated Protein Kinase Kinase 6; MKK 6; MKK6; MP2K6_HUMAN; PRKMK6; Protein kinase mitogen activated kinase 6; protein kinase, mitogen-activated, kinase 6 (MAP kinase kinase 6); SAPK kinase 3; SAPKK-3; SAPKK3; SKK3; Stress-activated protein kinase kinase 3;

Immunogens

Immunogen:

A synthesized peptide derived from human MKK3/MKK6 around the phosphorylation site of Ser189/Thr193.

Uniprot:

P46734(MP2K3_HUMAN) >>Visit HPA database.

P52564(MP2K6_HUMAN) >>Visit HPA database.

Gene(ID):
MAP2K6(5608) | MAP2K3(5606)
Expression:
P46734 MP2K3_HUMAN:

Abundant expression is seen in the skeletal muscle. It is also widely expressed in other tissues.

P52564 MP2K6_HUMAN:

Isoform 2 is only expressed in skeletal muscle. Isoform 1 is expressed in skeletal muscle, heart, and in lesser extent in liver or pancreas.

Sequence:
MESPASSQPASMPQSKGKSKRKKDLRISCMSKPPAPNPTPPRNLDSRTFITIGDRNFEVEADDLVTISELGRGAYGVVEKVRHAQSGTIMAVKRIRATVNSQEQKRLLMDLDINMRTVDCFYTVTFYGALFREGDVWICMELMDTSLDKFYRKVLDKNMTIPEDILGEIAVSIVRALEHLHSKLSVIHRDVKPSNVLINKEGHVKMCDFGISGYLVDSVAKTMDAGCKPYMAPERINPELNQKGYNVKSDVWSLGITMIEMAILRFPYESWGTPFQQLKQVVEEPSPQLPADRFSPEFVDFTAQCLRKNPAERMSYLELMEHPFFTLHKTKKTDIAAFVKEILGEDS

MSQSKGKKRNPGLKIPKEAFEQPQTSSTPPRDLDSKACISIGNQNFEVKADDLEPIMELGRGAYGVVEKMRHVPSGQIMAVKRIRATVNSQEQKRLLMDLDISMRTVDCPFTVTFYGALFREGDVWICMELMDTSLDKFYKQVIDKGQTIPEDILGKIAVSIVKALEHLHSKLSVIHRDVKPSNVLINALGQVKMCDFGISGYLVDSVAKTIDAGCKPYMAPERINPELNQKGYSVKSDIWSLGITMIELAILRFPYDSWGTPFQQLKQVVEEPSPQLPADKFSAEFVDFTSQCLKKNSKERPTYPELMQHPFFTLHESKGTDVASFVKLILGD

PTMs - P46734/P52564 As Substrate

Site PTM Type Enzyme
M1 Acetylation
S3 Phosphorylation
S15 Phosphorylation
S28 Phosphorylation
S31 Phosphorylation
K32 Ubiquitination
T39 Phosphorylation
K80 Ubiquitination
K93 Ubiquitination
S101 Phosphorylation
K105 Ubiquitination
K183 Ubiquitination
S189 Phosphorylation
K192 Ubiquitination
T193 Phosphorylation
K200 Ubiquitination
S212 Phosphorylation
Y214 Phosphorylation
S218 Phosphorylation Q99759 (MAP3K3) , P41279 (MAP3K8) , Q9UL54 (TAOK2) , Q16584 (MAP3K11) , Q7L7X3 (TAOK1) , Q99683 (MAP3K5) , P46734 (MAP2K3)
T222 Phosphorylation Q16584 (MAP3K11) , P46734 (MAP2K3)
Y230 Phosphorylation
K243 Ubiquitination
Y245 Phosphorylation
S249 Phosphorylation
S253 Phosphorylation
K279 Sumoylation
K279 Ubiquitination
K332 Ubiquitination
K340 Ubiquitination
Site PTM Type Enzyme
K5 Acetylation
K8 Acetylation
T25 Phosphorylation
S27 Phosphorylation
T28 Phosphorylation
K36 Ubiquitination
S40 Phosphorylation
Y64 Phosphorylation
K69 Ubiquitination
K82 Acetylation
K82 Ubiquitination
S90 Phosphorylation
K94 Ubiquitination
K141 Methylation
K141 Ubiquitination
K146 Ubiquitination
K172 Ubiquitination
S201 Phosphorylation
Y203 Phosphorylation
S207 Acetylation
S207 Phosphorylation P52564 (MAP2K6) , Q99683 (MAP3K5) , O43318 (MAP3K7) , P41279 (MAP3K8) , Q9UL54 (TAOK2)
T211 Acetylation
T211 Phosphorylation O43318 (MAP3K7) , Q13233 (MAP3K1) , Q9UL54 (TAOK2)
Y219 Phosphorylation P52564 (MAP2K6)
K232 Ubiquitination
K268 Sumoylation
K282 Ubiquitination
K296 Ubiquitination

PTMs - P46734/P52564 As Enzyme

Substrate Site Source
O15264 (MAPK13) T180 Uniprot
O15264 (MAPK13) Y182 Uniprot
P46734 (MAP2K3) S218 Uniprot
P46734 (MAP2K3) T222 Uniprot
P53778 (MAPK12) T183 Uniprot
P53778 (MAPK12) Y185 Uniprot
Q16539 (MAPK14) T180 Uniprot
Q16539 (MAPK14) Y182 Uniprot
Substrate Site Source
O15264-1 (MAPK13) T180 Uniprot
O15264 (MAPK13) Y182 Uniprot
P52564 (MAP2K6) S207 Uniprot
P52564 (MAP2K6) Y219 Uniprot
P53778 (MAPK12) T183 Uniprot
P53778 (MAPK12) Y185 Uniprot
Q14765 (STAT4) S721 Uniprot
Q15759 (MAPK11) T180 Uniprot
Q15759 (MAPK11) Y182 Uniprot
Q16539 (MAPK14) T180 Uniprot
Q16539-4 (MAPK14) Y182 Uniprot
Q9NQU5 (PAK6) T564 Uniprot
Q9NQU5 (PAK6) Y566 Uniprot
Q9ULV5 (HSF4) T471 Uniprot

Research Backgrounds

Function:

Dual specificity kinase. Is activated by cytokines and environmental stress in vivo. Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in the MAP kinase p38. Part of a signaling cascade that begins with the activation of the adrenergic receptor ADRA1B and leads to the activation of MAPK14.

PTMs:

Autophosphorylated. Phosphorylation on Ser-218 and Thr-222 by MAP kinase kinase kinases regulates positively the kinase activity. Phosphorylated by TAOK2.

Yersinia yopJ may acetylate Ser/Thr residues, preventing phosphorylation and activation, thus blocking the MAPK signaling pathway.

Tissue Specificity:

Abundant expression is seen in the skeletal muscle. It is also widely expressed in other tissues.

Subunit Structure:

Component of a signaling complex containing at least AKAP13, PKN1, MAPK14, ZAK and MAP2K3. Within this complex, AKAP13 interacts directly with PKN1, which in turn recruits MAPK14, MAP2K3 and ZAK. Binds to DYRK1B/MIRK and increases its kinase activity. Part of a complex with MAP3K3, RAC1 and CCM2 (By similarity). Interacts with ARRB1. Interacts with Yersinia yopJ.

Family&Domains:

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.

Function:

Dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. With MAP3K3/MKK3, catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in the MAP kinases p38 MAPK11, MAPK12, MAPK13 and MAPK14 and plays an important role in the regulation of cellular responses to cytokines and all kinds of stresses. Especially, MAP2K3/MKK3 and MAP2K6/MKK6 are both essential for the activation of MAPK11 and MAPK13 induced by environmental stress, whereas MAP2K6/MKK6 is the major MAPK11 activator in response to TNF. MAP2K6/MKK6 also phosphorylates and activates PAK6. The p38 MAP kinase signal transduction pathway leads to direct activation of transcription factors. Nuclear targets of p38 MAP kinase include the transcription factors ATF2 and ELK1. Within the p38 MAPK signal transduction pathway, MAP3K6/MKK6 mediates phosphorylation of STAT4 through MAPK14 activation, and is therefore required for STAT4 activation and STAT4-regulated gene expression in response to IL-12 stimulation. The pathway is also crucial for IL-6-induced SOCS3 expression and down-regulation of IL-6-mediated gene induction; and for IFNG-dependent gene transcription. Has a role in osteoclast differentiation through NF-kappa-B transactivation by TNFSF11, and in endochondral ossification and since SOX9 is another likely downstream target of the p38 MAPK pathway. MAP2K6/MKK6 mediates apoptotic cell death in thymocytes. Acts also as a regulator for melanocytes dendricity, through the modulation of Rho family GTPases.

PTMs:

Weakly autophosphorylated. Phosphorylated at Ser-207 and Thr-211 by the majority of M3Ks, such as MAP3K5/ASK1, MAP3K1/MEKK1, MAP3K2/MEKK2, MAP3K3/MEKK3, MAP3K4/MEKK4, MAP3K7/TAK1, MAP3K11/MLK3 and MAP3K17/TAOK2.

Acetylation of Ser-207 and Thr-211 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.

Subcellular Location:

Nucleus. Cytoplasm. Cytoplasm>Cytoskeleton.
Note: Binds to microtubules.

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

Isoform 2 is only expressed in skeletal muscle. Isoform 1 is expressed in skeletal muscle, heart, and in lesser extent in liver or pancreas.

Subunit Structure:

Dimer. Interacts with Yersinia yopJ. Interacts (via its D domain) with its substrates MAPK11, MAPK12, MAPK13 and MAPK14 (By similarity). Interacts (via its DVD domain) with MAP3Ks activators like MAP3K5/ASK1, MAP3K1/MEKK1, MAP3K2/MEKK2, MAP3K3/MEKK3, MAP3K4/MEKK4, MAP3K7/TAK1, MAP3K11/MLK3 and MAP3K17/TAOK2 (By similarity). Interacts with DCTN1. Interacts with EIF2AK2/PKR.

Family&Domains:

The DVD domain (residues 311-334) contains a conserved docking site and is found in the mammalian MAP kinase kinases (MAP2Ks). The DVD sites bind to their specific upstream MAP kinase kinase kinases (MAP3Ks) and are essential for activation.

The D domain (residues 4-19) contains a conserved docking site and is required for the binding to MAPK substrates.

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.

Research Fields

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

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

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

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

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Immune system > Toll-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.   (View pathway)

· Organismal Systems > Sensory system > Inflammatory mediator regulation of TRP channels.   (View pathway)

References

1). Structure-based discovery of 1H-indole-2-carboxamide derivatives as potent ASK1 inhibitors for potential treatment of ulcerative colitis. European Journal of Medicinal Chemistry, 2021 (PubMed: 33360793) [IF=6.7]

Application: WB    Species: mice    Sample: colon tissues

Fig. 6. Inhibitory effect of compounds 19 and 6 on activated ASK1-p38/JNK signaling and up-regulated inflammatory cytokines in colon tissues of DSS-induced UC mice. (A) Western blot analysis of the protein levels of ASK1-MKK3/6-p38 and ASK1-MKK4/7-JNK signaling pathways as well as corresponding phosphorylated proteins. GAPDH was used as the loading control protein. (B) Quantitative analysis of the ratios of phosphorylated proteins normalized to their unphosphorylated form. (C) The levels of inflammatory cytokines IL-1b, IL-6, TNF-a were detected using ELISA kits. Data are expressed as mean ± SEM (n ¼ 5e6 per group). Compared with the control group: # P < 0.05, ## P < 0.01, ### P < 0.001; Compared with the DSS þ Veh group: * P < 0.05, ** P < 0.01, *** P < 0.001.

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