Pin1 activity assay

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WebJun 1, 2024 · As the only known phosphorylation-dependent PPIase, Pin1 acts in tandem with Pro-directed kinases and phosphatases, which are conformation-specific, to control … Metrics - Identification of a potent and selective covalent Pin1 inhibitor - Nature WebNov 1, 2024 · Pin 1 Activity Assay Pin1 activity was determined as previously described ( 18, 30, 33) with some modifications. In short, neutrophils were resuspended in an ice cold lysis buffer containing 50 mM HEPES pH 7.5, 0.25% CHAPS, 100 mM NaCl, 1 mM β-glycerophosphate, 5 mM NaF and 1 mM EGTA, at 10 7 /ml, and lysed with several a 2 ml …

Essential role of Pin1 via STAT3 signalling and mitochondria …

WebMounting evidence has demonstrated that Pin1 is widely overexpressed and/or overactivated in cancer, exerting a critical influence on tumor initiation and progression … intech audio

PIN1 - an overview ScienceDirect Topics

WebStrikingly, while auxin transport during root initiation is mediated by the redundant activity of multiple PIN proteins, PIN1 is the dominant family member in cotyledon initiation … WebAug 29, 2024 · Pin1 is the only known peptidyl-prolyl cis–trans isomerase (PPIase) that specifically recognizes and isomerizes the phosphorylated Serine/Threonine-Proline … WebJun 2, 2014 · Specific activity is > 330 nmoles/min/mg, and is defined as the amount of enzyme that cleaves 1 µmole of suc-AAFP-pNA per minute at 25°C in Tris-Hcl pH8.0 using chymotrypsin. Activity Assay. 1. Prepare 180ul assay buffer into a suitable container: 133mM Tris-HCl, pH8.0, 5.5nM chymotrypsin, and 0.5ug of Pin1 recombinant protein. intech athens al

(A) PPIase activity assay using Pin1 as enzyme treatedw …

(PDF) Targeting Pin1 by inhibitor API-1 regulates ... - ResearchGate

WebPin1 activity can be measured in SNs or cell lysates by means of a protease-coupled, cis-to-trans isomerization assay ( 24, 32, 33 ). Conversion of the peptide substrate (Suc-Ala-Glu-Pro-Phe-pNA) from cis to trans permits cleavage of the C-terminal nitroaniline by chymotrypsin and detection at 390 nm. WebJan 30, 2024 · The general understanding of Pin1 in human cancer is that it promotes cancer occurrence and development by activating numerous oncogenic genes and … intech attendanceWebNational Center for Biotechnology Information intech athens alabama

"WebPin1 is a PPIase specific for pSer/pThr-Pro motifs; Uses recombinant human Pin 1 and substrate Succ-AEPF-pNA; Measures cis/trans isomerisation kinetics of substrate using … " - Pin1 activity assay

Pin1 activity assay

The role of Pin1 in the development and treatment of cancer

WebJun 30, 2013 · Pin1 SUMOylation inhibits its substrate binding and catalytic activity. A, molecular modeling using Discovery studio software and graphically depicted using PyMol. B, recombinant proteins were incubated with mitotic HeLa extracts followed by pulldown before subjecting to immunoblotting. WebPin1 consists of short N-terminal protein-protein interaction domain that allows enzyme to bind phosphoproteins and longer C-terminal izomerase domain. Pin1 has many biological …

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WebDec 20, 2024 · Pin1 is a peptidyl-prolyl isomerase responsible for isomerizing phosphorylated S/T-P motifs. Pin1 has two domains that each have a distinct ligand binding site, but only its PPIase domain has catalytic activity. WebEGCG inhibited Pin1 activity in a protease-coupled assay (IC 50 = 20 μM) and suppressed JNK signaling, cyclin D1 and Bcl-xL expression in ErbB2-transformed MEFs (Urusova et al. 2011 ). Tolerable doses of EGCG reduced the growth of ErbB2-transformed MEF xenografts in immune-deficient mice (Urusova et al. 2011 ).

WebJun 10, 2013 · Pin1 activity assay. g for 10 min (4°C). Pin1 activity was measured using equal amounts of artery cytoplasmic lysates and α-chymotrypsin using a synthetic tetrapeptide substrate Suc-Ala-Glu-Pro-Phe-pNa (Peptides International, Louisville, KY, USA). Absorption at 390 nM was measured using an Ultrospec 2000 spectrophotometer. WebFeb 1, 2024 · The trypsin activity assay kit can detect 10-100 mU (p-NA unit) trypsin. Trypsin activity assay protocol summary: - add samples, positive control and standards to wells - add chymotrypsin inhibitor to sample and positive control wells and incubate for 10 min at room temp - add reaction mix

WebFeb 1, 2024 · Pin1 (protein interacting with never in mitosis A-1) is a member of the peptidyl prolyl isomerase (PPIase) family, and catalyzes cis-trans isomerization of pThr/Ser-Pro … WebMay 19, 2024 · Pin1 consists of short N-terminal protein-protein interaction domain that allows enzyme to bind phosphoproteins and longer C …

WebThe SensoLyte® Green Pin1 Assay Kit uses a fluorogenic substrate pretreated to convert into cis isoform. Pin1 changes this substrate into the trans conformation that is readily …

WebJun 25, 2024 · The optical density of each well in the assay, which in proportional to both the LDH activity and percentage of necrotic cells, was measured at a wavelength of 490 or 450 nm with a microplate reader. The percentage of necrotic cell death was calculated from four independent experiments by measuring the optical density of the treated group minus ... jobs white house tnWebOriginal magnification, 40. (g) Luciferase assay of lysates prepared from 293 cells at 36 h after transfection with pEF1-lacZ and pUAS-Luc and expressing Pin1 and Gal4-IRF3 with or without the RIG ... in tech automation in rochester nhWebDec 12, 2024 · PIN1 is a peptidyl-prolyl isomerase that catalyzes the cis / trans isomerization of peptide bonds between proline and phosphorylated serine/threonine residues. By … intech automation incWebMar 27, 2024 · Multifaceted approaches were taken to determine the inhibition profiles of these compounds against Pin1 activity in vitro and in vivo, including an isomerization assay, surface plasmon resonance (SPR) technology, virtual docking, MTT proliferation assay, western blotting, cell cycle analysis, apoptosis analysis, immunofluorescence analysis ... intech automotive engineering s de rl de cvWebMar 27, 2024 · Small molecules targeting Pin1 as potent anticancer drugs Front Pharmacol. 2024 Mar 27;14:1073037. doi: 10.3389/fphar.2024.1073037. eCollection 2024. Authors … intech automation puneWebJul 8, 2024 · Using co-immunoprecipitation assay, the interaction between HBx and Pin1 was further confirmed in a cellular context of HepG2 cells. ... Pin1 can enhance the transcriptional activity of cyclin D1 ... jobs whitehorseWebSep 21, 2009 · Pin1 activity assay. Pin1 assay was performed as described (g for 10 minutes (4°C). PPIase activity was measured using equal amounts of parathyroid cytoplasmic lysates and α-chymotrypsin using a synthetic tetrapeptide substrate Suc-Ala-Glu-Pro-Phe-pNa (Peptides International). Absorption at 390 nM was measured using an … jobs whiterock