AFfirm™ VASP Mouse Monoclonal Antibody - #BF8131

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製品: VASP Mouse Monoclonal Antibody
カタログ: BF8131
タンパク質の説明: Mouse monoclonal antibody to VASP
アプリケーション: WB IHC
反応性: Human, Mouse
予測: Pig, Bovine, Sheep, Dog
分子量: 46kDa,50kDa; 40kD(Calculated).
ユニプロット: P50552

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製品説明

ソース:
Mouse
アプリケーション:
WB 1:500-1:3000, IHC 1:50-1:200
*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,Mouse
クローナリティ:
Monoclonal [AFfirm8131]
特異性:
VASP Antibody detects endogenous levels of total VASP.
コンジュゲート:
Unconjugated.
精製:
Affinity-chromatography.
保存:
Mouse IgG1 in phosphate buffered saline (without Mg2+ and Ca2+), 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

Vasodilator stimulated phosphoprotein; Vasodilator-stimulated phosphoprotein; VASP; VASP_HUMAN;

免疫原

免疫原:
Uniprot:

P50552(VASP_HUMAN) >>Visit HPA database.

遺伝子(ID):
VASP(7408)
発現特異性:
P50552 VASP_HUMAN:

Highly expressed in platelets.

タンパク質の説明:
Vasodilator-stimulated phosphoprotein (VASP) is a member of the Ena-VASP protein family. Ena-VASP family members contain an EHV1 N-terminal domain that binds proteins containing E/DFPPPPXD/E motifs and targets Ena-VASP proteins to focal adhesions.
タンパク質配列:
MSETVICSSRATVMLYDDGNKRWLPAGTGPQAFSRVQIYHNPTANSFRVVGRKMQPDQQVVINCAIVRGVKYNQATPNFHQWRDARQVWGLNFGSKEDAAQFAAGMASALEALEGGGPPPPPALPTWSVPNGPSPEEVEQQKRQQPGPSEHIERRVSNAGGPPAPPAGGPPPPPGPPPPPGPPPPPGLPPSGVPAAAHGAGGGPPPAPPLPAAQGPGGGGAGAPGLAAAIAGAKLRKVSKQEEASGGPTAPKAESGRSGGGGLMEEMNAMLARRRKATQVGEKTPKDESANQEEPEARVPAQSESVRRPWEKNSTTLPRMKSSSSVTTSETQPCTPSSSDYSDLQRVKQELLEEVKKELQKVKEEIIEAFVQELRKRGSP

PTMs - P50552 基板として

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
S8 Phosphorylation
Y16 Phosphorylation
Y39 Phosphorylation Q9UM73 (ALK)
T43 Phosphorylation
S46 Phosphorylation
R68 Methylation
K71 Ubiquitination
S134 Phosphorylation
S157 Phosphorylation P17252 (PRKCA) , P17612 (PRKACA) , Q15139 (PRKD1) , Q13976 (PRKG1) , Q9BZL6 (PRKD2)
S239 Phosphorylation Q13976 (PRKG1) , Q13237 (PRKG2) , P17612 (PRKACA) , Q9BZL6 (PRKD2)
T249 Phosphorylation
K252 Acetylation
S258 Phosphorylation
T278 Phosphorylation Q15418 (RPS6KA1) , P54646 (PRKAA2) , P17612 (PRKACA) , Q13131 (PRKAA1) , Q13976 (PRKG1)
K283 Acetylation
T284 Phosphorylation
K286 Sumoylation
S289 Phosphorylation
S305 Phosphorylation
K312 Ubiquitination
S314 Phosphorylation
T315 Phosphorylation
T316 Phosphorylation
K321 Methylation
K321 Ubiquitination
S322 Phosphorylation P54646 (PRKAA2) , Q9BZL6 (PRKD2) , Q15139 (PRKD1)
S323 Phosphorylation
S324 Phosphorylation
S325 Phosphorylation
T327 Phosphorylation
T328 Phosphorylation
T331 Phosphorylation
T335 Phosphorylation
S337 Phosphorylation
Y341 Phosphorylation
K348 Ubiquitination
K363 Ubiquitination

研究背景

機能:

Ena/VASP proteins are actin-associated proteins involved in a range of processes dependent on cytoskeleton remodeling and cell polarity such as axon guidance, lamellipodial and filopodial dynamics, platelet activation and cell migration. VASP promotes actin filament elongation. It protects the barbed end of growing actin filaments against capping and increases the rate of actin polymerization in the presence of capping protein. VASP stimulates actin filament elongation by promoting the transfer of profilin-bound actin monomers onto the barbed end of growing actin filaments. Plays a role in actin-based mobility of Listeria monocytogenes in host cells. Regulates actin dynamics in platelets and plays an important role in regulating platelet aggregation.

PTMs:

Major substrate for cAMP-dependent (PKA) and cGMP-dependent protein kinase (PKG) in platelets. The preferred site for PKA is Ser-157, the preferred site for PKG/PRKG1, Ser-239. In ADP-activated platelets, phosphorylation by PKA or PKG on Ser-157 leads to fibrinogen receptor inhibition. Phosphorylation on Thr-278 requires prior phosphorylation on Ser-157 and Ser-239. In response to phorbol ester (PMA) stimulation, phosphorylated by PKC/PRKCA. In response to thrombin, phosphorylated by both PKC and ROCK1. Phosphorylation at Thr-278 by AMPK does not require prior phosphorylation at Ser-157 or Ser-239. Phosphorylation at Ser-157 by PKA is required for localization to the tight junctions in epithelial cells. Phosphorylation modulates F-actin binding, actin filament elongation and platelet activation. Phosphorylation at Ser-322 by AMPK also alters actin filament binding. Carbon monoxide (CO) promotes phosphorylation at Ser-157, while nitric oxide (NO) promotes phosphorylation at Ser-157, but also at Ser-239. Response to NO and CO is blunted in platelets from diabetic patients, and VASP is not phosphorylated efficiently at Ser-157 and Ser-239.

細胞の位置付け:

Cytoplasm. Cytoplasm>Cytoskeleton. Cell junction>Focal adhesion. Cell junction>Tight junction. Cell projection>Lamellipodium membrane. Cell projection>Filopodium membrane.
Note: Targeted to stress fibers and focal adhesions through interaction with a number of proteins including MRL family members. Localizes to the plasma membrane in protruding lamellipodia and filopodial tips. Stimulation by thrombin or PMA, also translocates VASP to focal adhesions. Localized along the sides of actin filaments throughout the peripheral cytoplasm under basal conditions. In pre-apoptotic cells, colocalizes with MEFV in large specks (pyroptosomes).

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:

Highly expressed in platelets.

サブユニット構造:

Homotetramer. Interacts with PFN1, PFN2, LPP, ACTN1 and ACTG1. Interacts, via the EVH1 domain, with the Pro-rich regions of ZYX. This interaction is important for targeting to focal adhesions and the formation of actin-rich structures at the apical surface of cells. Interacts, via the EVH1 domain, with the Pro-rich domain of Listeria monocytogenes actA. Interacts with APBB1IP. Interacts, via the Pro-rich domain, with the C-terminal SH3 domain of DNMBP (By similarity). Interacts weakly with MEFV.

タンパク質ファミリー:

The EVH2 domain is comprised of 3 regions. Block A is a thymosin-like domain required for G-actin binding. The KLKR motif within this block is essential for the G-actin binding and for actin polymerization. Block B is required for F-actin binding and subcellular location, and Block C for tetramerization.

The WH1 domain mediates interaction with XIRP1.

Belongs to the Ena/VASP family.

研究領域

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

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

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.   (View pathway)

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

参考文献

1). Pharmacological inhibition of Septins with Forchlorfenuron attenuates thrombus formation in experimental thrombotic mice models with modulating multiple signaling pathways in platelets. Journal of advanced research, 2024 (PubMed: 39111626) [IF=10.7]

Application: WB    Species: human    Sample:

Fig. 6. The effect of FCF on human platelet to agonists stimulation. Washed human platelets were pre-incubated with different doses of FCF (25, 50, and/or 100 µM) at 37 °C for 10 min, subsequently subjected into platelets aggregation, western blotting, adhesion and clot retraction assays in response to stimulation with thrombin (0.01 U/mL), collagen (2 µg/mL) and/or ADP (20 µM). (A) Bars graph showing the quantified inhibition rates of platelet aggregation (n = 3 each group). (B) Representative western blotting showing phosphorylation levels of PI3K, AKT, GSK3β, ERK1/2, p-p38, JNK, VASP (S157) and VASP (S239) (n = 3 each group). (C) Human platelets were subjected on FN matrix for one hour in presence of 0.01 U/mL thrombin, and IF images showing platelet morphological alternation and intracellular CD62P expression (Red, F-actin; Green, CD62P). Bar graphs quantified (D) adherent platelet number, (E) averaged platelet area and (F) CD62P MFI values (n = 3 each group). (G) The representative images showing human platelets colt retract progression and (H) the curve plots summarized relative colt retraction (n = 3 each group). (I) human platelets supernatants from aggregation assay (FCF, 100 µM) were harvested for ATP releases by luciferin/luciferase reagents and the data are presented as the relative percentage to solvent (n = 3 each group). The graphs summarize the data from at least three times independent experiments, data are shown as the mean ± S.E.M., and analyzed using a One-way ANOVA or Unpaired student’s t-test, and *p < 0.05, **p < 0.01, and ***p < 0.001 versus solvent (Sol.). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

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