I was wondering if TEV protease would work on reverse seuqnece? A typical cleavage recognition site for TEV protease is ENLYFQ/G with cleavage between Q and G. Would TEV protease work if the sequence is reversed (G\QFYLNE)?
No. It is specific for the sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) and cleaves between the Gln and Gly/Ser residues.
However, this paper reports that the P1' specificity (the residue at the C-terminal end of the cleavage site) is more relaxed than that.
https://www.sciencedirect.com/science/article/abs/pii/S0006291X02005740
This paper also investigates the substrate specificity of TEV, but offers no indication that any residue other than Q is allowed at P1 (the residue at the N-terminal end of the cleavage site).
Article Substrate Profiling of Tobacco Etch Virus Protease Using a N...
To rephrase the question, how does the TEV protease distinguishe between the cleavable site sequence ---ENLYGFQ/SG--- versus ---GS/QFGYLNE---?
Based on the stated specificity, the enzyme would not recognize the second sequence.
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