Interaction of free porphyrins and metalloporphyrins with mouse ferrochelatase. A model for the active site of ferrochelatase.
The ability of purified mouse ferrochelatase (protoheme ferro-lyase, EC 4.99.1.1) to bind and catalytically utilize a variety of porphyrins has been examined. In all, the kd, Km or Ki values for eleven different porphyrins, the Ki values for four metalloporphyrins and the kd values for two metalloporphyrins were determined. The data obtained demonstrate that mouse ferrochelatase binds a wide variety of porphyrins and metalloporphyrins with kd values ranging from 6 nM for N-methylprotoporphyrin to 1.08 microM for coproporphyrin III. However, the enzyme shows a degree of catalytic specificity for the substituents at the 2.4 positions and utilizes only proto-, hemato-, meso-, deutero-, 2,4-monohydroxy-ethylmonovinyl- and 2,4-monohydroxymethylmonovinyl deuteroporphyrin as substrates. The data show that the magnitude of the kd is not an accurate indicator of the ability of the porphyrin to serve as a substrate or inhibitor and, with the exception of N-methylprotoporphyrin, the size of the kd is several orders of magnitude less than that of the Km or Ki. Of the metalloporphyrins examined (Fe, Co, Zn and Sn) all inhibited ferrochelatase at micromolar concentrations, although tin protoporphyrin was the least effective. These data are discussed in terms of an active site model for mammalian ferrochelatase.
Dailey
H A
HA
Department of Microbiology, University of Georgia, Athens 30602.
Jones
C S
CS
Karr
S W
SW
D000818:Animals; D001665:Binding Sites; D003035:Cobalt / Q000494:pharmacology; D005294:Ferrochelatase / Q000378:metabolism*; D007501:Iron / Q000494:pharmacology; D007700:Kinetics; D008190:Lyases / Q000378:metabolism*; D008665:Metalloporphyrins / Q000378:metabolism; D008745:Methylation; D051379:Mice; D015394:Molecular Structure; D011166:Porphyrins / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D008665:Metalloporphyrins; D011166:Porphyrins; D003035:Cobalt; D007501:Iron; D008190:Lyases; D005294:Ferrochelatase