Identification of lipase folding intermediates through chemically induced denaturation using fluorescence.

<p>A) Chemically induced unfolding of native LipA from <i>B. glumae</i> is displayed by plotting the fluorescence emission intensity at 330 nm (in arbitrary units) against the denaturant concentration at two different incubation times (1 h incubation represented with open circles, 16 h incubation represented with black filled circles). GuHCl-induced denaturation of LipA_n upon 16 h incubation reveals the existence of an unfolding intermediate. B) Intrinsic protein fluorescence and ANS binding study for different lipase folding conformations: native lipase (LipA_n; black circles), near-native intermediate (LipA_i; red circles), molten globule-like conformation in 1.2 M guHCl (LipA_g; green inverse triangles), unfolded lipase in presence of 6 M guHCl (LipA_u; yellow triangles).