Recombinant Human Thiopurine S-Methyltransferase (TPMT) Protein (GST)

Name: Recombinant Human Thiopurine S-Methyltransferase (TPMT) Protein (GST)
Brand: Beta LifeScience
SKU: BLC-08535P
Availability: InStock

Greater than 90% as determined by SDS-PAGE.

Based on the SEQUEST from database of E.coli host and target protein, the LC-MS/MS Analysis result of this product could indicate that this peptide derived from E.coli-expressed Homo sapiens (Human) TPMT.

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Product Overview

Description	Recombinant Human Thiopurine S-Methyltransferase (TPMT) Protein (GST) is produced by our E.coli expression system. This is a protein fragment.
Purity	Greater than 90% as determined by SDS-PAGE.
Uniprotkb	P51580
Target Symbol	TPMT
Synonyms	HGNC:12014; S adenosyl L methionine thiopurine S methyltransferase; Thiopurine methyltransferase; Thiopurine S methyltransferase; Thiopurine S-methyltransferase; TPMT; TPMT_HUMAN
Species	Homo sapiens (Human)
Expression System	E.coli
Tag	N-GST
Target Protein Sequence	TRTSLDIEEYSDTEVQKNQVLTLEEWQDKWVNGKTAFHQEQGHQLLKKHLDTFLKGKSGLRVFFPLCGKAVEMKWFADRGHSVVGVEISELGIQEFFTEQNLSYSEEPITEIPGTKVFKSSSGNISLYCCSIFDLPRTNIGKFDMIWDRGALVAINPGDRKCYADTMFSLLGKKFQYLLCVLSYDPTKHPGPPFYVPHAEIERLFGKICNIRCLEKVDAFEERHKSWGIDCLFEKLYLLTE
Expression Range	4-244aa
Protein Length	Partial
Mol. Weight	54.7kDa
Research Area	Metabolism
Form	Liquid or Lyophilized powder
Buffer	Liquid form: default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. Lyophilized powder form: the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Storage	1. Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. 2. Avoid repeated freeze-thaw cycles. 3. Store working aliquots at 4°C for up to one week. 4. In general, protein in liquid form is stable for up to 6 months at -20°C/-80°C. Protein in lyophilized powder form is stable for up to 12 months at -20°C/-80°C.
Notes	Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Target Details

Target Function	Catalyzes the S-methylation of thiopurine drugs such as 6-mercaptopurine (also called mercaptopurine, 6-MP or its brand name Purinethol) and 6-thioguanine (also called tioguanine or 6-TG) using S-adenosyl-L-methionine as the methyl donor. TPMT activity modulates the cytotoxic effects of thiopurine prodrugs. A natural substrate for this enzyme has yet to be identified.
Subcellular Location	Cytoplasm.
Protein Families	Class I-like SAM-binding methyltransferase superfamily, TPMT family
Database References	HGNC: 12014 OMIM: 187680 KEGG: hsa:7172 STRING: 9606.ENSP00000312304 UniGene: PMID: 29491687 In Mexican patients, mutant alleles were detected in 6.2 and 5.2% of systemic lupus erythematosus and rheumatoid arthritis cases, respectively. PMID: 29264794 TPMT3C affects TPMT activity in Chinese patients with neuromyelitis optica spectrum disorders. PMID: 29191122 Patients also developed infection who mostly (71%) needed hospitalization. None of the studied G238C, G460A and A719G TPMT variant alleles were detected. Infections and febrile neutropenia were common causes of 6-PM dose modification and interruption. PMID: 29321348 ANCA associated vasculitis (AAV) patients treated with cyclophosphamide induction and azathioprine maintenance therapy were included and followed for 60 months. TPMT genotype and activity were not independent predictors of relapse, and could not predict leukopenia or other adverse effects from azathioprine. PMID: 29630648 Results show no association between TPMT or COMT single-nucleotide polymorphisms and cisplatin-induced ototoxicity. PMID: 28445188 Of the 14 patients intolerant to thiopurines, NGS identified deleterious TPMT variants in 5 individuals whereas the biochemical test identified 8 individuals as intolerant (sensitivity 35.7% and 57.14%; specificity 93.75% and 50% respectively). SKAT-O test identified a significant association between MOCOS gene and TPMT activity (p = 0.0015), not previously reported. PMID: 27703193 A statistically significant correlation between the presence of the G460A mutation and the development of leucopenia after the administration of thiopurines was observed. PMID: 28857898 genetic association studies in cohort in Denmark: Data suggest that standard immunosuppression treatment failure in patients with autoimmune hepatitis is not associated with thiopurine S-methyltransferase genetic variants or HLA-DRB103 genotype in the population studied. (HLA-DRB1 = major histocompatibility complex class II DR beta 1) PMID: 28374975 This article reviews the thiopurine pharmacogenetics and the methods applied in common practice to evaluate patient's TPMT status. [review] PMID: 28552060 study aimed to determine the relationship between thiopurine metabolite levels and therapeutic response, and to investigate the association of NUDT15, TPMT, and thiopurine metabolites with leukopenia in patients with CD PMID: 29206869 TPMT3C is overrepresented in acute lymphoblastic leukemia (ALL) cases in comparison with non-ALL group PMID: 28476189 TMPT polymorphisms are associated with 6-mercaptopurine intolerance in children treated for acute lymphoblastic leukemia. PMID: 27577869 The frequency of TPMT mutation in a homogeneous Sicilian cohort of patients with inflammatory bowel disease, autoimmune and hematological disorders was low. TPMT genotyping is not a sensitive tool for predicting thiopurine-induced leukopenia. PMID: 27665263 meta-analysis of three independent GWAS of TPMT activity; only genetic variants mapping to chromosome 6, including the TPMT gene region, were significantly associated with TPMT activity;finding is consistent with TPMT genotype being the primary determinant of TPMT activity PMID: 27770449 In a GWAS, TPMT activity in patients behaves as a monogenic trait, further bolstering the utility of TPMT genetic testing in the clinic PMID: 27564568 Review/Meta-analysis: thiopurine S-methyltransferase testing for averting drug toxicity. PMID: 27217052 A point mutation in the thiopurine S-methyltransferase gene that led to exon 5 deletion in the transcribed mRNA. PMID: 26974142 The aims of this research were to investigate the association of TPMT phenotypes with MDR1 genotypes. PMID: 27453281 The aim of the study was to investigate frequencies of TPMT and ITPA polymorphisms in Lithuanian inflammatory bowel disease patients and analyze their association with azathioprine-related adverse events. PMID: 26674571 With respect to TPMT, the variants TPMT2 and TPMT3A were not implicated in genetic susceptibility to childhood acute lymphoblastic leukemia PMID: 26845729 structure-function relationships of TPMT PMID: 26410243 The TPMT promoter region may serve as a pharmacogenomic biomarker when introducing thiopurine therapy PMID: 26411491 The most frequently occurring nonfunctional TPMT allele in Croatian population is TPMT3A. Variant genotypes were statistically significantly more common in Crohn's disease subgroup than in ulcerative colitis subgroup. PMID: 27333713 Association of TPMT polymorphisms with overall azathioprine-induced adverse drug reactions, bone marrow toxicity and gastric intolerance, but not with hepatotoxicity (meta-analysis). PMID: 26633017 association of TPMT polymorphisms with overall thiopurine-induced adverse drug reactions PMID: 25799415 Identification of TPMT variants and subsequent dose reduction reduces hematologic events during thiopurine treatment of inflammatory bowel disease. PMID: 26072396 TPMT3c mutant allele was associated with azathioprine side effects (leukopenia, alopecia) in Chinese systemic lupus erythematosus patients. PMID: 24322830 refinements in risk stratification and treatment have reduced the influence of TPMT genotype on treatment outcome in a contemporary protocol. PMID: 25940902 We report on the presence of the TPMT3C and 3A mutant alleles in the Libyan population. Therefore, monitoring the patients to be treated with doses of thiopurine drugs for TPMT variants is worthwhile to avoid the development of severe myelosuppression. PMID: 25819542 this paper shows that the influence of TPMT and COMT on the development of cisplatin-induced hearing loss may be less important than previously suggested. PMID: 25551397 Results show complete sequence-based screening study evaluating all TPMT variants in Asian populations some of them may ne of relevance in Korean population. PMID: 25564374 distribution of the most frequent variants of TPMT gene was similar in a healthy population and patients with inflammatory bowel disease PMID: 23581716 The prevalence of TPMT genotypes was high among Brazilian patients. Variants genes 2 and 3C may be associated with azathioprine pancreatic toxicity in a IBD southeastern Brazilian population PMID: 24696613 Data suggest risk of myelotoxicity of high-dose methotrexate during methotrexate/6-mercaptopurine maintenance chemotherapy of childhood acute leukemia is not associated with heterozygosity in thiopurine methyltransferase in Nordic population studied. PMID: 25347948 TMPT genotyping appears an important tool to further optimize 6-MP treatment design in Chilean patients with ALL PMID: 24774509 suggest that germline genetic variation in TPMT and MTHFR do not significantly alter SOS risk in patients exposed to thioguanine PMID: 24737678 TPMT37 introduces a premature stop codon at position 216, resulting in loss of the last 29 amino acid residues from the C terminal of the TPMT protein. PMID: 24710034 plays a key role in the metabolism of azathioprine/6- mercaptopurine (6-MP). Mutations in the enzyme lead to generation of excess thioguanine, which causes neutropenia via suppression of neutrophils. PMID: 23996738 Activity measurement performed at diagnosis provides clinicians with information on immediate pharmacokinetic-related adverse events and/or hypermetabolism PMID: 24643197 Heart transplantation recipients with TPMT genetic variant alleles who are treated with AZA develop acute rejection earlier, more frequently, and of greater severity. PMID: 24121523 The study focused on explaining how a locally occurred TPMT A167G substitution propagated through hydrogen bonds alteration to induce structural modification which affects both thiopurine and S-adenosylmethionine receptors. PMID: 23025308 This study shows the prevalence of TPMT genetic polymorphisms in population of Slovak IBD patients PMID: 23731044 TPMT3A and the TPMT3C, which cause the largest decrease in enzyme activity, were both variant alleles detected in the Tunisian population PMID: 23553048 In childhood acute lymphoblastic leukaemia, TPMT activity should not be used to predict heterozygosity particularly in blood samples obtained at disease diagnosis PMID: 23252716 differential contribution of the enzyme TPMT to the cytotoxicity of the two thiopurines is probably due to its role in formation of the meTIMP, the cytotoxic methylated metabolite of 6-MP PMID: 23811272 A predictive model combining variants in TPMT, ABCC3, and COMT with clinical variables significantly improved the prediction of hearing-loss development as compared with using clinical risk factors alone PMID: 23588304 our results indicated that TPMT or COMT genetic variation was not related to cisplatin ototoxicity in children with cancer and did not influence cisplatin-induced hearing damage in laboratory models PMID: 23820299 A high frequency and diversity of variant TPMT genotypes was found in this series with predominance of the TPMT3B allele PMID: 23844534 In a study of Italian myasthenia gravis patients treated with azathioprine, a new TPMT haplotype, TPMT 3E, was observed only in association with intolerance. PMID: 23400745

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

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