Search PubMed⌕ Search

Biomedical subjects

Patricia Ann Mabrouk

Publications and source records attributed to Patricia Ann Mabrouk.

7 recordsLinked to original sources
PubMed2003

Proton NMR study of chemically modified horse heart ferricytochrome c confirms the presence of histidine and lysine-ligated conformers in 30% acetonitrile solution.

Comparison of the 1H NMR spectra for guanidinated ferricyt c and chloro(terpyridine)platinum(II)-modified ferricyt c in 30% acetonitrile (ACN) solution with that for ferricyt c in 30% ACN is reported. The absence of the heme methyl proton resonances characteristic of the IV*-form (Lys-ligated) in the NMR spectrum of guanidinated ferricyt c in 30% ACN solution confirms that a lysine-ligated form of ferricyt c is produced in 30% ACN solution. The absence of the 8-methyl heme proton resonance of the V*-form in the NMR spectrum of chloro(terpyridine)platinum(II)-modified ferricyt c in 30% ACN solution demonstrates that a bis-His-ligated form of ferricyt c is produced in 30% ACN, not a hydroxide ligated form, as previously proposed. The revised assignment for the V* form of ferricyt c in mixed media explains differences between the exchange network we previously reported for ferricyt c in 30% ACN [Protein Sci. 10 (2001) 2291] as versus that reported by Dopner et al. at high pH [J. Am. Chem. Soc. 120 (1998) 11246]. Lys- and His-ligated forms are known to be produced in the presence of denaturants in protein folding studies of ferricyt c. Consequently, the exchange network between these non-native forms of ferricyt c in 30% ACN may have biological relevance for ferricyt c folding.

Acetonitriles↗
PubMed2003

Structure-function relationship of reduced cytochrome c probed by complete solution structure determination in 30% acetonitrile/water solution.

The complete solution structure of ferrocytochrome c in 30% acetonitrile/70% water has been determined using high-field 1D and 2D (1)H NMR methods and deposited in the Protein Data Bank with codes 1LC1 and 1LC2. This is the first time a complete solution protein structure has been determined for a protein in nonaqueous media. Ferrocyt c retains a native protein secondary structure (five alpha-helices and two omega loops) in 30% acetonitrile. H18 and M80 residues are the axial heme ligands, as in aqueous solution. Residues believed to be axial heme ligands in the alkaline-like conformers of ferricyt c, specifically H33 and K72, are positioned close to the heme iron. The orientations of both heme propionates are markedly different in 30% acetonitrile/70% water. Comparative structural analysis of reduced cyt c in 30% acetonitrile/70% water solution with cyt c in different environments has given new insight into the cyt c folding mechanism, the electron transfer pathway, and cell apoptosis.

Acetonitriles↗
PubMed2002

Spectroscopic and electrochemical studies of horse myoglobin in dimethyl sulfoxide.

This paper reports the first report of rapid, reversible direct electron transfer between a redox protein, specifically, horse myoglobin, and a solid electrode substrate in nonaqueous media and the spectroscopic (UV-vis, fluorescence, and resonance Raman) characterization of the relevant redox forms of myoglobin (Mb) in dimethyl sulfoxide (DMSO). In DMSO, the heme active site of metmyoglobin (metMb) appears to remain six-coordinate high-spin, binding water weakly. Changes in the UV-fluorescence spectra for metMb in DMSO indicate that the protein secondary structure has been perturbed and suggest that helix A has moved away from the heme. UV-vis and RR spectra for deoxyMb in DMSO suggest that the heme iron is six-coordinate low-spin, most likely coordinating DMSO. Addition of CO to deoxyMb in DMSO produces a single, photostable six-coordinate CO adduct. UV-vis and RR for Mb-CO in DMSO are consistent with a six-coordinate low-spin heme iron binding His93 weakly, if at all. The polarity of the distal heme pocket is comparable to that of the closed form of horse Mb-CO in aqueous solution, pH 7. Direct electron transfer between horse Mb and Au in DMSO solution was investigated by cyclic voltammetry. Mb exhibits stable and well-defined electrochemical responses that do not appear to be affected by the water content (1.3-7.5%). The electrochemical characteristics are consistent with a one-electron, quasi-reversible, diffusion-controlled charge transfer process at Au. E degrees for horse Mb in DMSO at Au is -0.241+/-0.005 V vs. NHE. The formal heterogeneous electron transfer rate constant, calculated from delta E(p) at 20 mV/s, is 1.7+/-0.5 x 10(-4) cm/s. The rate, which is unaffected by the presence of 1.3-7.5% water, is competitive with that previously reported for horse Mb in aqueous solution.

Animals↗
PubMedDate not supplied

Raman evidence that the lyoprotectant poly(ethylene glycol) does not restore nativity to the heme active site of horseradish peroxidase suspended in organic solvents.

Resonance Raman spectroscopy was used to interrogate the heme active site of horseradish peroxidase (HRP) lyophilized in the presence and absence of the lyoprotectant poly(ethylene glycol) (PEG; FW 5000; 0-80% w/w) suspended in acetone, chloroform, or acetonitrile. In aqueous solution, Fe(3+)HRP is characterized by a five-coordinate high-spin (5-c HS) heme system. The structure of the heme-active site of HRP in all solvents is perturbed by co-lyophilization of HRP with PEG. Heme active site structural changes are consistent with coordination of water in the distal axial coordination site of the ferric heme iron and disruption of the hydrogen-bond network when the protein is lyophilized in the presence of PEG (>or=60% w/w) in all of the solvent systems studied. Similar active site structural changes were previously observed for HRP in benzene and attributed to a change in the reaction mechanism for HRP in benzene. (Mabrouk, P. A.; Spiro, T. G. J. Am. Chem. Soc. 1998, 120, 10303-10309.) Thus, PEG is proposed to increase the catalytic activity of HRP in nonaqueous media by locking the heme active site into a structure that functions through an alternative catalytic pathway in nonaqueous media.

Binding Sites↗