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KMID : 0383119550030060059
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Journal of Aerospace Medicine
1955 Volume.3 No. 6 p.59 ~ p.76
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Paper Electrophoretic and Paper Chromatographic Studies of the Blood Proteins
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Abstract
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It has been known that fetal hemoglobin is different from adult hemoglobin. However, there has been many controversy among authors- on the heterogeneity of adult hemoglobin. Betke and White claimed that there are two components in adult hemoglobin as the results of alkaline denaturation reaction and peroxidase activity test. After that Itano doubted the above results because the alkaline denaturation reaction by itself can contribute little to the elucidation of this problem. But, thereafter, Vat Fossan reported that adult hemoglobin could be separated into 3 components by aluminum oxide column chromatography using oxyhemoglobin. If there should exist more than one components, it might bring forward many interesting problems in such cases as anemia´s, hypoxia and so forth just like the serum protein fractions. Accordingly, first of all, it is necessary to elucidate whether the hemoglobin, molecule is homogeneous or heterogeneous.
The author adopted the paper electrophoretic and paper chromatographic procedures in addition to the Van Fossan´s column chromatography. Moreover, the author used the most stable cyanomethemoglobin instead of oxyhemoglobin, because the latter undergoes easily autooxidation evens ii the, air. For the paper electrophoresis, applied 103-403´ volts and Walpole´s acetate and Michaelis ammonia and ver9nal buffers of various-pH of which ionic strengthen range from. 0.5 to 6:1 Toyo No.2, No, 51, Whatman No. 4 and Eaton Dikeman No. 613 filter papers were employed. For the paper- chromatography same kinds of paper and buffer Were employed, an&, ascending method was performed. For the column chromatography adopted almost same procedures as Van Fossan´s,
Experimental results can be summarized as follows.
By paper electrophoresis the author could not recognize fractions´ in normal adult hemoglobin, regardless of the kinds, ionic strengthes: and pH of the-:buffers and voltage applied, Then, if there should be more than two components, each imaginary components may be different in other properties other than their isoelectric points molecular weights and solubilities, because by Svedberg´s ultracentrifuge one cannot recognize more than one component and Itano´s experiment on solubility was also counter-evidential. If these ma mar hemoglobin components are adsorptive properties, column chromatographic separation might be possible as Van Fossan´s report But the author´s result using cyanomethemoglobin instead of oxyhemoglobin were negative. In author´s new approach using paper chromatography by buffer solutions, sometimes there appeared two components in pH, near 2.9 but these apparent fractions was identified to be pseudofractions by two Dimension development. The cause of such apparent separation could be explained easily by Gibbs theory.
At last, the author discussed on the so called pseudofractions of hemoglobin and general applicability of the other authors methods, and concluded that normal adult hemoglobin molecule is homogeneous single substance.
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