Recombinant Human Adenosylhomocysteinase (AHCY)

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Code CSB-EP001474HU
MSDS
Size $224
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Product Details

Purity
Greater than 90% as determined by SDS-PAGE.
Target Names
AHCY
Uniprot No.
Research Area
Metabolism
Alternative Names
Adenosylhomocysteinase; AdoHcyase; ahcY; S adenosyl L homocysteine hydrolase; S adenosylhomocysteine hydrolase; S-adenosyl-L-homocysteine hydrolase; SAHH; SAHH_HUMAN
Species
Homo sapiens (Human)
Source
E.coli
Expression Region
2-432aa
Target Protein Sequence
SDKLPYKVADIGLAAWGRKALDIAENEMPGLMRMRERYSASKPLKGARIAGCLHMTVETAVLIETLVTLGAEVQWSSCNIFSTQDHAAAAIAKAGIPVYAWKGETDEEYLWCIEQTLYFKDGPLNMILDDGGDLTNLIHTKYPQLLPGIRGISEETTTGVHNLYKMMANGILKVPAINVNDSVTKSKFDNLYGCRESLIDGIKRATDVMIAGKVAVVAGYGDVGKGCAQALRGFGARVIITEIDPINALQAAMEGYEVTTMDEACQEGNIFVTTTGCIDIILGRHFEQMKDDAIVCNIGHFDVEIDVKWLNENAVEKVNIKPQVDRYRLKNGRRIILLAEGRLVNLGCAMGHPSFVMSNSFTNQVMAQIELWTHPDKYPVGVHFLPKKLDEAVAEAHLGKLNVKLTKLTEKQAQYLGMSCDGPFKPDHYRY
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Mol. Weight
63.6kDa
Protein Length
Full Length of Mature Protein
Tag Info
N-terminal 6xHis-SUMO-tagged
Form
Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
3-7 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA
Please contact us to get it.
Description

Insertion of the gene encoding the Human AHCY protein (2-432aa) into a plasmid vector results in the formation of recombinant plasmid, which is then introduced into e.coli cells. Positive e.coli cells are selected relying on their ability to survive in the presence of a specific antibiotic. The e.coli cells containing the recombinant plasmid are cultured under conditions conducive to the expression of the gene of interest. A N-terminal 6xHis-SUMO tag is attached to the protein. Following expression, the recombinant Human AHCY protein is isolated and purified from the cell lysate through affinity purification. The resultant recombinant Human AHCY protein is analyzed using denaturing SDS-PAGE, revealing a purity greater than 90%.

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Target Background

Function
Adenosylhomocysteine is a competitive inhibitor of S-adenosyl-L-methionine-dependent methyl transferase reactions; therefore adenosylhomocysteinase may play a key role in the control of methylations via regulation of the intracellular concentration of adenosylhomocysteine.
Gene References into Functions
  1. Results uncover a H19/SAHH circuit involving gene-methylation alterations by carcinogen BaP. PMID: 29772428
  2. We investigated previously assumed interaction with AHCY-like-1 protein (AHCYL1), a paralog of AHCY. Indeed, significant interaction between both proteins exists. Additionally, silencing AHCYL1 leads to moderate inhibition of nuclear export of endogenous AHCY. PMID: 28647132
  3. We have validated the vectors and confirmed self-association of AHCY, AHCYL1, and galectin-3. In a high-throughput BiFC screen, we identified new AHCY interaction partners: galectin-3 and PUS7L. We also describe additional steps in protein interaction analysis, applied for AHCY-galectin-3 interaction PMID: 27455993
  4. In order to enable the development of small molecule AHCY inhibitors as targeted cancer therapeutics we developed an assay based on a RapidFire high-throughput mass spectrometry detection system, which allows the direct measurement of AHCY enzymatic activity. PMID: 28533090
  5. SAH hydrolase deficiency can remain asymptomatic in childhood, and the disorder can be associated with early onset hepatocellular carcinoma. PMID: 26527160
  6. H19 knockdown activates SAHH, leading to increased DNMT3B-mediated methylation of an lncRNA-encoding gene Nctc1 within the Igf2-H19-Nctc1 locus. PMID: 26687445
  7. S-adenosylhomocysteine hydrolase is regulated by lysine acetylation PMID: 25248746
  8. SAHH can promote apoptosis, inhibit migration and adhesion of ESCC cells suggesting that it may be involved in carcinogenesis of the esophagus. PMID: 24430301
  9. A fluorescence-based assay for the measurement of S-adenosylhomocysteine hydrolase activity in biological samples. PMID: 23079506
  10. The simulations of ligand-induced transition revealed that the signal of intrasubunit closure dynamics is transmitted to form intersubunit contacts, which in turn invoke a precise alignment of active site. PMID: 22023331
  11. Five active site residues (E156, N181, K186, D190, N191) of AdoHcy hydrolase have been individually mutated to alanine and each engineered enzyme characterized with respect to its redox partial reaction and elimination/addition partial reaction. PMID: 12069606
  12. a discussion of its hydrolytic activity (review) PMID: 12369977
  13. Maintenance of ionizable active-site residues in catalytically suitable protonation states in closed forms of placental AHCY may be assisted by a water chain, stabilized by Asp182, that can import and export protons from and to the environment. PMID: 12590576
  14. AdoHcy is involved in adenosine-induced apoptosis by altering gene expression. PMID: 17097637
  15. R38W and G123R amino-acid exchanges did not bring about major changes to the catalytic rates. However, circular dichroism analysis showed that both polymorphisms effect the thermal stability. PMID: 17164794
  16. SAHH, which is diffuse in the cytoplasm of nonmotile Dictyostelium amoebae and human neutrophils, concentrates with F-actin in pseudopods at the front of motile, chemotaxing cells PMID: 17172447
  17. In the case of Hs-SAHH, the slow-binding phase terminates in micromolar affinity, but over a period of hours, the dissociation rate constant decreases until the final equilibrium affinity is in the nanomolar range. PMID: 17447732
  18. We found clinically relevant levels of Hcy (0-500 microM) induced elevation of SAH, declination of SAM and SAM/SAH ratio and reduced expression of SAHH and MBD2, but increased activity of DNMT3a and DNMT3b affecting DNA methylation PMID: 17688412
  19. The mechanism of action of copper on this enzyme sugges a regulative role for copper on the intracellular activity. PMID: 17892301
  20. consider changes in charge and the sterical incompatibility in mutant p.A89V protein as main reason for enzyme malfunction with AdoHcyase deficiency as consequence PMID: 18211827
  21. SAHH from Homo sapiens (Hs-SAHH) and from the parasite Trypanosoma cruzi (Tc-SAHH) are very similar in structure and catalytic properties but differ in the kinetics and thermodynamics of association and dissociation of the cofactor NAD (+) PMID: 18393535
  22. Streptococcal pyrogenic exotoxin B (SPE B) potentially causes immunosuppression by cleaving human S-adenosylhomocysteine hydrolase (AdoHcyase). PMID: 18522500
  23. increased plasma concentration in patients with idiopathic thrombocytopenic purpura PMID: 18683034
  24. SAHH mRNA was lost in 50% of tumor tissues from 206 patients with different kinds of tumors in comparison with normal tissue counterparts. Moreover, SAHH protein was also affected in some colon cancers PMID: 18713839
  25. These data show that 2-5-fold enhanced AdoHcyase activity is well tolerated by the cell, while greatly enhanced AdoHcyase activity results in adenosine-induced apoptosis. PMID: 18769049
  26. S-adenosylhomocysteine hydrolase PMID: 19619139

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Involvement in disease
Hypermethioninemia with S-adenosylhomocysteine hydrolase deficiency (HMAHCHD)
Subcellular Location
Cytoplasm. Melanosome. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Protein Families
Adenosylhomocysteinase family
Database Links

HGNC: 343

OMIM: 180960

KEGG: hsa:191

STRING: 9606.ENSP00000217426

UniGene: Hs.388004

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