2 μg NleL run on 4-12% SDS-PAGE gel under reducing conditions

Active, Escherichia coli O157:H7, Recombinant NleL

Enzyme

Product Code: TE3-020

Verified Applications: In vitro, recombinant NleL accepts activated ubiquitin from E2 conjugating enzyme UBE2L3 in a reaction that also requires Ubiquitin Activating Enzyme 1 (an E1). The charged E3 demonstrates strong autoubiquitylation and generation of unanchored poly-ubiquitin in the absence of added substrates. Appropriate enzyme concentrations are specific to the application.

Activity: Verified in Poly-ubiquitin Chain Synthesis Assay

Purity: 95%

Concentration: 15 – 20 μM

For Research Use Only (RUO)

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Protein Name: NleL

Predicted Molecular Mass: 70 kDa kDa

Background and Additional Info

Background: NleL is a type III secretion system effector protein produced by pathogenic Escherichia coli strains. It functions as an E3 ubiquitin ligase, modulating host cell signaling pathways by ubiquitinating host proteins, thereby promoting infection. NleL comprises an N-terminal β-helix formed by pentapeptide repeats (residues 1–250) and a C-terminal bilobed bacterial HECT (bHECT) catalytic domain. key motifs include the catalytic Cys387 and an E2-binding patch. NleL positions itself downstream of host E2 enzymes, directly catalyzing ubiquitin transfer to host substrates or building unanchored K6- and K48-linked poly-ubiquitin chains. References Franklin, T.G., et al. (2023) Mol Cell 83:4538–4554. PMID 38091999 Piscatelli, H., et al., (2011) PLoS ONE 6:e19331. PMID 21541301 Lin, D. Y-W., et al., (2012) PNAS 109:1925-1930. PMID 22308380 Hospenthal, M. K., et al., (2013) Nat. Struct. Mol. Biol. 20:555-565. PMID 23563141 Sheng, X., et al., (2017 ) PLoS Pathog 13:e1006534. PMID 28753655

Shipping: The product is shipped with dry ice. Upon receipt, store it immediately at the temperature recommended below.

Storage and Stability: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Aliquot and store ≤ -70°C (stable for 24 months from date of receipt).

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Alternate Names: NleL (Non-LEE encoded effector L)

Protein Sequence: GPLGSQGRACLSKAELTADLIWLSANRTGEESAEELNYSGCDLSGLSLVGLNLSSVNFSGAVLDDTDLRMSDLSQAVLENCSFKNSILNECNFCYANLSNCIIRALFENSNFSNSNLKNASFKGSSYIQYPPILNEADLTGAIIIPGMVLSGAILGDVKELFSEKSNTINLGGCYIDLSDIQENILSVLDNYTKSNKSILLTMNTSDDKYNHDKVRAAEELIKKISLDELAAFRPYVKMSLADSFSIHPYLNNANIQQWLEPICDDFFDTIMSWFNNSIMMYMENGSLLQAGMYFERHPGAMVSYNSSFIQIVMNGSRRDGMQERFRELYEVYLKNEKVYPVTQQSDFGLCDGSGKPDWDDDSDLAYNWVLLSSQDDGMAMMCSLSHMVDMLSPNTSTNWMSFFLYKDGEVQNTFGYSLSNLFSESFPIFSIPYHKAFSQNFVSGILDILISDNELKERFIEALNSNKSDYKMIADDQQRKLACVWNPFLDGWELNAQHVDMIMGSHVLKDMPLRKQAEILFCLGGVFCKYSSSDMFGTEYDSPEILRRYANGLIEQAYKTDPQVFGSVYYYNDILDRLQGRNNVFTCTAVLTDMLTEHAKESFPEIFSLYYPVAWR