Reconstitution hemoproteins

Farrell H, Correia MA. Structural and functional reconstitution of hepatic cytochrome P450 in vivo. Reversal of allylisopropylacetamide-mediated destruction of the hemoprotein by exogenous heme. J Biol Chem 1980 255 10128-10133. 

Table 1 gives the names and abbreviations used for the hemoproteins. The reconstituted cyanoferrimyoglobins, where protoheme IX was replaced by deuteroheme IX or mesoheme IX, will be referred to as DeutMbmCN and MesoMbinCN. For mixed state hemoglobins the states of the individual subunits will be indicated. For example in Hb l1 jSspCN) the a-chains would be in the deoxy-form, and the /9-chains in the cyanoferri-form. 

Boddupalli, S. S., Oster T., Estabrook, R. W., and Peterson, J. A. 1992b. Reconstitution of the Fatty-Acid Hydroxylation Function of Cytochrome-P-450bm-3 Utilizing Its Individual Recombinant Hemoprotein and Flavoprotein Domains. J. Biol. Chem., 267,10375-10380. 

Iron porphines constitute the active site of hemoproteins and as such are among the most important and most widely studied series of metal complexes. The syntheses of iron protoporphyrin (heme) and its derivatives are important for the reconstitution of hemoproteins (particularly when one wishes to incorporate 57Fe for Mossbauer studies), as well as for model studies. In addition to the naturally occurring porphyrins, iron complexes of both 5,10,15,20-tetraphenyl-21//,23//-porphine (H2tpp) and octaethyl-21//,23//-porphine (H2oep) are widely used in model studies of the natural systems. 

S. typhimurium enzyme (S94), which appears to be essentially identical to the E. coli sulfite reductase, are in agreement with the above results. Thus mutants lacking the flavoprotein or the hemoprotein component of the enzyme and containing only the appropriate partial activities have been obtained and the respective partial enzymes isolated. The absorption spectra of sulfite reductase preparations from the wild type and from these mutants are shown in Fig. 46, and the proposed structure for the two components of the wild-type enzyme is shown in Fig. 47. Reconstitution of NADPH-sulfite reductase by recombination of the flavoprotein... 

An alternative approach to the preparation of photoenzymes by the reconstitution of apo-proteins includes the application of metalloprotoporphyrins such as Zn(II)-protoporphyrin IX (Zn-P), which exhibits photophysical electron-transfer properties for the reconstitution of apo-proteins derived from hemoproteins. Func-... 

Rats were treated with vehicle (control), phenobarbital (PB), or 3-methylcholanthrene (3-MC). Cytochrome P450, lipid, and reductase fractions were prepared and reconstituted. The reductase and lipid fractions were prepared from PB-treated rats. No hydroxylation activity was detected when hemoprotein was omitted from the reaction mixture. In Experiment 1, benzo[a]pyrene metabolism was measured by formation of fluorescent phenolic metabolites, and benzphetamine metabolism was measured by the rate of benzphetamine-dependent NADPH oxidation. In Experiment 2, the metabolism of pentobarbital, benzo[a]pyrene, and chlorcyclizine was measured by product formation. Experiment 1 was taken from Ref. (53) and Experiment 2 was taken from Ref. (55). 

Willner, I., Heleg-Shalilai, V., Katz, E., Rau, H.K., and Haehnel, W. (1999) Integration of a reconstituted de novo synthesized hemoprotein and native metalloproteins with electrode supports for bioelectronic and bioelectrocatalytic applications. Journal of the American Chemical Society, 121, 6455-6468. 

Extensive research efforts were directed in recent years towards the synthesis of de novo proteins. ° Specifically, the reconstitution of de novo proteins with metal prophyrin attracted interest as a biomimetic approach to tailor synthetic hemoproteins. ... 

Interaction of the functionalized-monolaycr electrode with Fe(III)-protoporphyrin IX, (15), resulted in the reconstitution of the protein with the two heme units. The surface coverage of the reconstituted de novo hemoprotein was found to be 2.5x10 mole cm Assuming that the protein footprint area is ca. 25 A, the saturated surface coverage of the de novo protein corresponds to 3.5x 10" mole cm and thus the experimental value... 

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