Proton NMR study of the interaction of tin(IV) protoporphyrin IX monomers and dimers with apomyoglobin.
Events during the reconstitution of apomyoglobin to form the holoprotein were probed by porphyrin-metal substitution. Thus interactions between tin(IV) protoporphyrin IX (SnPP) and equine apomyoglobin (apoEqMb), and between tin(IV) protoporphyrin IX dimers [(SnPP)2] and apoEqMb, were observed by 1H NMR and optical absorbance spectroscopic techniques. The chief advantages of using SnPP are that products and intermediates can easily be related to SnPP.EqMb which has been studied [Deeb, R.S., & Peyton, D.H. (1991) J. Biol. Chem. 266, 3728-3733] and that at least one step during reconstitution is slowed considerably as compared to heme. Reactions of apoEqMb with SnPP and (SnPP)2 produce different intermediates, although the final product, SnPP.EqMb, is the same for each. An intermediate observed for reaction of SnPP with apoEqMb at pH 10 is in exchange with free SnPP, with the observed rate constant koff approximately 1 s-1. meso-Proton resonances were assigned for this intermediate by correlation to SnPP resonances via chemical exchange. The intermediate observed for reaction of (SnPP)2 with apoEqMb at pH 7.5 is heterogeneous. The reaction of either SnPP or (SnPP)2 with apoEqMb at neutral pH produces another species which may be the alternate porphyrin-insertion isomer arising from a 180 degree rotation about the alpha, gamma-meso axis of the porphyrin. Although optical absorbance spectroscopy of the Soret region shows evidence for each reaction, only in combination with 1H NMR are the various processes assigned.
Deeb
R S
RS
Chemistry Department, Portland State University, Oregon 97207-0751.
Peyton
D H
DH
D001059:Apoproteins / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008665:Metalloporphyrins / Q000737:chemistry*; D009211:Myoglobin / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D011522:Protons; D011524:Protoporphyrins / Q000737:chemistry*; D013055:Spectrophotometry, Infrared
D001059:Apoproteins; D008665:Metalloporphyrins; D009211:Myoglobin; D011522:Protons; D011524:Protoporphyrins; C010857:apomyoglobin; C032628:tin protoporphyrin IX