Myoglobin Oxygen Binding Curves Determined by Phosphorescence Quenching of Palladium Porphyrin

A completely optical method was developed for evaluating myoglobin-oxygen saturation. Solution oxygen concentrations were computed from measurements of phosphorescence decay of a soluble palladium porphyrin according to the Stern-Volmer quenching relationship. Visible absorption spectra were recorded of myoglobin solutions progressively deoxygenated by bacterial aerobic metabolism. Myoglobin oxygenation curves were obtained from the spectra by two full-spectrum procedures, singular value decomposition (SVD) and curve fitting, and by the traditional dual-wavelength isosbestic method. The calculated P50 for horse heart myoglobin at 24 ± 1°C was 1.2 Torr by SVD, 1.2 Torr by curve fitting, and 1.4 Torr by dual-wavelength spectroscopy. These results compare favorably with a value of 1.1 Torr estimated from the work of Rossi-Fanelli and Antonini for human myoglobin. The Hill coefficient, theoretically 1.00 for myoglobin, was experimentally determined to be 0.96 by SVD and 0.95 by curve fitting of the spectral data matrix.

[1]  S Nioka,et al.  Time-resolved spectroscopy of hemoglobin and myoglobin in resting and ischemic muscle. , 1988, Analytical biochemistry.

[2]  J. Vanderkooi,et al.  Phosphorimeters for analysis of decay profiles and real time monitoring of exponential decay and oxygen concentrations. , 1988, Analytical Biochemistry.

[3]  W. Rumsey,et al.  Imaging of phosphorescence: a novel method for measuring oxygen distribution in perfused tissue. , 1988, Science.

[4]  W. Rumsey,et al.  The oxygen dependence of mitochondrial oxidative phosphorylation measured by a new optical method for measuring oxygen concentration. , 1988, The Journal of biological chemistry.

[5]  J. Vanderkooi,et al.  An optical method for measurement of dioxygen concentration based upon quenching of phosphorescence. , 1987, The Journal of biological chemistry.

[6]  B. Hedlund,et al.  Application of a coulometric oxygen sensor to the determination of hemoglobin--oxygen binding isotherms. , 1977, Analytical biochemistry.

[7]  F. Jöbsis,et al.  Pyrenebutyric acid as an optical oxygen probe in the intact cerebral cortex. , 1976, Journal of applied physiology.

[8]  T. Yonetani,et al.  Absorption and luminescence of yttrium and lanthanide octaethylporphin complexes , 1976 .

[9]  M. Tsutsui,et al.  Unusual metalloporphyrins. XVI. Preparation and purification of tetrasodium meso-tetra(p-sulfophenyl)porphine. Easy procedure , 1973 .

[10]  J. Knopp,et al.  Intracellular measurement of oxygen by quenching of fluorescence of pyrenebutyric acid. , 1972, Biochimica et biophysica acta.

[11]  G. Weber,et al.  Oxygen quenching of pyrenebutyric acid fluorescence in water. A dynamic probe of the microenvironment. , 1970, Biochemistry.

[12]  L. Sklar,et al.  A mechanistic study of the synthesis of meso‐tetraphenylporphin , 1968 .

[13]  E. Antonini,et al.  Studies on the oxygen and carbon monoxide equilibria of human myoglobin. , 1958, Archives of biochemistry and biophysics.

[14]  F. Roughton,et al.  The equilibrium between oxygen and sheep haemoglobin at very low percentage saturations , 1951, The Journal of physiology.

[15]  C. Barlow,et al.  Metabolic Spectroscopy for Monitoring Electromagnetic Medical Techniques , 1990 .

[16]  W. Rumsey,et al.  Oxygen distribution in isolated perfused liver observed by phosphorescence imaging. , 1989, Advances in experimental medicine and biology.

[17]  J. Bonaventura,et al.  Thin-layer methods for determination of oxygen binding curves of hemoglobin solutions and blood. , 1981, Methods in enzymology.

[18]  M. Hitchman,et al.  Measurement of Dissolved Oxygen , 1978 .

[19]  E. Antonini.  INTERRELATIONSHIP BETWEEN STRUCTURE AND FUNCTION IN HEMOGLOBIN AND MYOGLOBIN. , 1965, Physiological reviews.