Non-ionic detergent affects the conformation of a functionally active mutant of Bcl-X(L).

We found that a mutant, Bcl-X(L)(F131V), which was previously reported to have impaired binding capacity, can bind to Bax almost as strongly as wild-type Bcl-X(L). In the absence of detergent, the Bcl-X(L)(F131V) mutant adopts the same conformation as wild-type Bcl-X(L), as determined by circular dichroism spectroscopy and size-exclusion chromatography. However, non-ionic detergent induces a conformational change in the Bcl-X(L)(F131V) mutant and causes it to lose Bax-binding capacity. Wild-type Bcl-X(L), on the other hand, is more resistant to detergent-induced effects and retains its ability to bind Bax in the presence of detergent. Since it has been shown that the Bcl-X(L)(F131V) mutant has nearly the same anti-apoptotic activity as wild-type Bcl-X(L), it would be likely that the Bcl-X(L)(F131V) mutant can adopt the wild-type conformation, rather than the detergent-induced conformational state and can bind to Bax in vivo. Therefore, our data demonstrated that non-ionic detergent can have unpredicted effects on protein conformation, differential effects on wild-type and mutant Bcl-X(L) proteins in this case and may cause complications in the interpretation of in vitro binding studies.