Just a curious question:
- What will happen if i remove most protein sequence that encoded for non-binding site of Restriction Enzyme to make a new enzyme, can the new restriction enzyme still work?
As it seems with about everything in protein science, the answer is it depends on the protein. Many proteins will lose activity if they are truncated; however, I've worked with GPCR's that were truncated down to the extracellular portion only and they showed consistent kinetic results. Antibodies have been cut into pretty tiny chunks (scFv's) to create some drugs like Pexelizumab. Some proteins can be hacked down quite a bit, some can't be at all.
The active site usually consists of amino acids from multiple different regions of the enzyme. So if you remove or replace parts of the protein chain then the structure (folding) of the enzyme will be most probably different and the active site will cease to exist. On the other hand in lucky cases replacing a few amino acids can increase the catalytic activity.