Just a quick note: there is a thermal shift-like methd that is still htp
and allows one to measure unfolding of membrane proteins etc. The method
relies on a cys-reactive leuco dye that becomes fluorescent (or colored)
after it reacts with an sh. As long as your protein has at least one buried
SH t
Unfortunately, their isn't a "general rule" for good versus bad
nucleophiles. However, their are some key classical examples, which are
reviewed extensively in enzymology textbooks. Remember, that a nucleophile
is dependent on it's pKa value, which depends on the surrounding residues.
Thus, two dif
Biochemistry and enzymology texts are a good place to start. In general,
stronger bases are better nucleophiles (it doesn't correspond exactly) but
you also have to consider the population of the deprotonated species at the
relevant pH, and the role of neighboring residues in altering 'native' pKa
