HEPARIN-INDUCED CONFORMATIONAL CHANGE AND ACTIVATION OF MUCUS PROTEINASE-INHIBITOR

Low molecular mass heparin (5.1 kDa) forms a tight complex with mucus proteinase inhibitor, the physiologic neutrophil elastase inhibitor of the upper respiratory tract. This binding strongly enhances the intrinsic fluorescence of the inhibitor and the rate of neutrophil elastase inhibitor association. One mole of this heparin fragment binds 1 mol of inhibitor with a K(d) of 50 nM. From the variation of K(d) with ionic strength, it is inferred that (i) 85% of the heparin-inhibitor binding energy is due to electrostatic interactions, (ii) about seven ionic interactions are involved in heparin-inhibitor binding, and (iii) about one-third of the ionized charges of heparin and inhibitor are involved in the complex. Heparin 4-fold increases the very low quantum yield of Trp30, the single tryptophan residue of the inhibitor, blue-shifts its maximum emission wavelength by 6 nm, decreases the acrylamide quenching rate constant by a factor of 4, and increases the mean intensity weighted lifetime by a factor of 2.5. These important spectroscopic changes evidence a heparin-induced conformational change of the inhibitor which buries Trp30 in a very hydrophobic environment. Heparin accelerates the inhibition of elastase in a concentration-dependent manner. When both enzyme and inhibitor are saturated by the polymer, the second-order association rate constant is 7.7 X 10(7) M-1 s-1, a value that is 27-fold higher than that measured with the free partners. This finding may have important physiologic and therapeutic bearing.