UbiQ Probes, activity-based DUB probes
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UbiQ Probes, activity-based DUB probes Target identification and validation
단백질 탈유비퀴틴화에 관련된 효소의 crystallization, identification, validation에 유용합니다. 프로브는 많은 DUB가 가지는 활성 부위 시스테인 잔기에 반응하도록 설계되어 있으며, C 말단에는 반응기가 결합하고, N 말단에 검출용 태그가 부가되어 있습니다. 반응기는 친전자체(electrophile)로 작용하고 친핵 (nucleophile) 활성 부위인 시스테인 잔기와 반응합니다.
Activity-based probes for deubiquitinases (DUBs) are based on ubiquitin with a C-terminal warhead designed to react with the active site cysteine residue present in most DUBs. UbiQ offers a selection of activity-based probes (ABPs) with various N-terminal tags. They enable the researcher to crystallize, identify and validate dozens of DUBs and ubiquitin like proteases enzymes as potential targets involved in protein deubiquitination.
Three generations of DUB probes: 1. 1st generation of DUB probes target the S1 pocket of a DUB and have contributed greatly to our understanding of DUBs. 2. 2nd generation of DUB probes target the S1 and S1’ pockets. These are di-ubiquitin-based probes with an electrophilic group between the two ubiquitins. 3. 3rd generation of DUB probes target the S1 and S2 pocket and are useful tools to identify DUBs that rely on S2 site interactions to fine tune their activity.
Activity-based DUB probes
UbiQ’s wide selection of activity-based DUB probes consists of various electrophiles, substrates and N- terminal tags. In addition there is the option to order tailor-made activity based probes.
Electrophiles UbiQ offers three different electrophiles: propargylamide (PA or Prg), vinylmethyl ester (VME) and vinylsulfone (VS). All are irriversibe DUB inhibitors and together these target all major DUB families.
* A recently discovered DUB family see References 7 ** According to a proteomics study the PA probes also target DUBs of the Machado-Josephin Domain family.1
N-terminal tags
Applications • Structural studies, target crystallization • Phenotypic protein profiling on cell lysates for target validation • Chemical proteomics for target identification (i.e., pull down, purification, western blotting, MS)
Experience • In addition to various activity-based probes, UbiQ has developed more than15 custom probes for a range of customers • Numerous successfully crystallized DUBs using UbiQ’s activity-based probes, including: USP4, UCH-L5, and the viral ovarian tumor DUB (vOTU), and Ub-PA probes. • Target validation and identification of DUBs in infectious diseases (University Glasgow / GSK)
References 1 (a) Ekkebus et al. J Am Chem Soc 2013, 135, 2867. (b) Sommer et al. Bioorg Med Chem 2013, 21, 2511. 2 (a) de Jong et al. ChemBioChem 2012, 13, 2251. (b) Altun et al. Chem Biol 2011, 18, 1401. 3 (a) Mulder & El Oualid et al. ChemBioChem 2014, 15, 946. (b) El Oualid et al. Angew. Chem. Int. Ed. 2010, 49, 10149. 4 (a) Misaghi et al. J. Biol. Chem. 2005, 280, 1512. (b) Galardy et al. Methods in Enzymology 2005, 399, 120. 5 (a) Borodovsky et al. Chemistry and Biology 2002, 9, 1149. (b) Borodovsky et al. EMBO J. 2001, 20, 5187. 6 (a) Clerici, M., et al. Nat.Commun. 2014, 5, 5399. (b) Sahtoe, D.D. et al.Mol.Cell 2015, 57, 887 7 (a) Rehman, A., et al. Mol Cell. 2016, 63, 1. (b) Bekes, M. et al. Mol.Cell 2016, 62, 572 For a complete list of references please we refer to the product group overview document.
Brochure (PDF)
Ordering informations
1st generation DUB probes: target the S1 pocket
* Size ; 50ug
2nd generation DUB probes: target the S1 and S1’ pockets
* Size ; 25ug
3rd generation DUB probes: target the S1 and S2 pockets
* Size ; 25ug
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▣ 관련 페이지 ; UbiQ Bio BV
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