FAST EVENTS IN PROTEIN-FOLDING - RELAXATION DYNAMICS AND STRUCTURE OFTHE I-FORM OF APOMYOGLOBIN

The fast relaxation dynamics of the acid destabilized I form of apomyo globin (pH 3, 0.15 M NaCl: apoMb-I) following a laser-induced tempera ture-jump have been probed using time-resolved infrared spectroscopy. Only a fast, single exponential phase is observed (bleach centered at v = 1633 cm(-1) and transient absorbance at 1666 cm(-1)) with relaxati on times of 38 ns at 30 degrees C and 36 ns at 57 degrees C; no additi onal slow (microsecond) phase is observed as previously found in the n ative form of apomyoglobin, Folding times of approximately 66 ns are d erived from the observed rates based on a simple two-state model. The equilibrium melting of the 1633 cm(-1) component shows noncooperative linear behavior over the temperature range studied (10-60 degrees C). The low amide I' frequency, the fast relaxation dynamics, and the nonc ooperative melting behavior are characteristic of isolated solvated he lix. The analysis of the amide-I' band reveals another major component at 1650 cm(-1) assigned to native-like structure stabilized by tertia ry contacts involving the AGH core, which does not show dynamic or sta tic melting under our conditions. ApoMb-I has generally been taken to be a ''molten globule'' species, The present results indicate a hetero geneous structure consisting of separate regions of native-like unit(s ), solvated helices, and disordered coil, excluding a homogeneous molt en globule as a model for apoMb-I. From the current studies and other results, a detailed model of the folding of apomyoglobin is presented.