[Isolation and properties of myoglobin from murine skeletal muscle].
Myoglobin (Mb) was isolated from skeletal muscle of JCL-ICR mice by heat denaturation-gel filtration combined with ion exchange chromatography or chromatofocusing by which isoelectric point of the main component was estimated as 7.63 +/- 0.09 (20 degrees C). The Mb was homogeneous by gel electrophoretic and ultracentrifugal analysis. The molecular weight by sedimentation equilibrium was 1.80 X 10(4) and essentially identical with the values by the iron analysis (1.82 X 10(4)) and amino acid composition (1.78 X 10(4)) in which one residue of cysteine was found per molecule. The spectroscopic properties of deoxy-, oxy-, carboxy- and ferri-derivatives of the protein were determined in ultraviolet, Soret and visible regions. The pK' of acid-alkaline transition of the ferri-form was estimated as 8.57 +/- 0.30 (30 degrees C) from the pH-dependent spectral changes. The oxygen equilibrium studies revealed complete absence of such allosteric properties as heme-heme interaction, anion effect and Bohr effect. Oxygen tension for the half-oxygenation (P50) was 0.69 +/- 0.06 Torr (20 degrees C) and its temperature-dependent change gave the delta H degrees of -14.1 kcal/mole.
Nakatani
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A
Second Department of Physiology, Nara Medical University.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis*; D009211:Myoglobin / Q000032:analysis / Q000302:isolation & purification*; D010100:Oxygen; D013056:Spectrophotometry, Ultraviolet; D014461:Ultracentrifugation
D000596:Amino Acids; D009211:Myoglobin; D010100:Oxygen