Difference Spectroscopic Titration and formation of β-hematin.

<p>Panels A and B show difference absorption spectra of heme binding to BNT1 and BNTM respectively. Panel C shows heme-binding curves of BNT1 and BNTM generated from the difference absorption spectra by plotting A₄₁₅ vs the moles of heme/mole of peptide template. Horizontal lines were drawn connecting the Y axis to the plateau region of the binding curves. At the point of intersection, vertical arrows were drawn to obtain the values of heme/peptide.Panel D shows comparative kinetics of heme binding to BNT1/BNTM (at heme: peptide molar ratio of 4) studied by difference spectroscopy. Data shows intensity of Soret peak as a function of time. Panel E shows comparative kinetics of β-hematin formation in presence of BNT1/BNTM.