A heme binding site on myelin basic protein: characterization, location, and significance.
Myelin basic protein (MBP), an extrinsic membrane protein from the myelin sheath, binds dicyanohemin. The binding generates absorption bands in the Soret region and quenches the fluorescence emitted by the sole tryptophan residue. The absorption titration curves in the Soret demonstrate that the binding is stoichiometric, one heme per protein, with a large value of the extinction coefficient (8 X 10(4) M-1 cm-1 at 420 nm). Fluorescence quenching titration curves indicate an identical stoichiometry and a low quenching efficiency of 20%. From the heme titration curve the association constant between dicyanohemin and MBP is estimated to be greater than or equal to 10 nM-1 in 50 mM 4-morpholinepropanesulfonic acid buffer, pH 7.0, at 20 degrees C. Digestion of MBP by Staphylococcus aureus V8 protease yields a peptide (38-118) whose heme binding properties are identical to those of MBP. In contrast, peptides obtained by digestion of MBP with cathepsin D do not exhibit any specific binding of dicyanohemin. The cleavage of the Phe-Phe (42-43) bond appears to be critical in this respect. A comparison of the sequence immediately preceding, including these residues with a probable heme binding site of a mitochondrial cytochrome b, reveals a high degree of homology. The possible significance of heme binding is discussed.
D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D001665:Binding Sites; D002402:Cathepsin D; D002403:Cathepsins; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D002942:Circular Dichroism; D006418:Heme / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000378:metabolism*; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D012997:Solvents; D013050:Spectrometry, Fluorescence
D000596:Amino Acids; D004676:Myelin Basic Protein; D010446:Peptide Fragments; D012997:Solvents; D006418:Heme; D002403:Cathepsins; D010447:Peptide Hydrolases; D002402:Cathepsin D