E. coli O157:H7 NleL Catalytic Domain Protein, CF Summary
Escherichia coli O157:H7 recombinant NleL Catalytic Domain is a bacterial HECT class Ubiquitin Ligases (E3) that catalyzes the formation of mixed K6 and K48-linked unanchored polyubiquitin chains. Reaction conditions will need to be optimized for each specific application. We recommend an initial E. coli O157:H7 recombinant NleL concentration of 0.1-0.5 μM.
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
X mg/ml (X μM) in 50 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol (v/v), 1 mM TCEP
|Shipping||The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
NleL (Non-Lee-encoded effector Ligase, also known as espX7) is an effector protein translocated into host cells via the type III secretion system of Escherichia coli O157:H7, a widespread pathogen causing severe foodborne diseases. Although the biological relevance of NleL is presently unknown, recent studies clearly demonstrate that this protein is functionally related to the HECT (Homologous to E6AP Carboxy Terminus) class E3 Ubiquitin Ligases of higher eukaryotes. NleL shows no primary sequence homology to any eukaryotic proteins, but biochemical and structural studies revealed both N-terminal and C-terminal lobes connected by a flexible linker, and a catalytic cysteine that accepts ubiquitin from E2 carrier proteins, both hallmarks of HECT domain ligases. NleL catalyzes the formation of mixed K6 and K48-linked unanchored polyubiquitin chains. This recombinant protein contains amino acid residues 170-782 (UniProt Q8X5G6 numbering).
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