The Effects of Urea on the Dissociation of Lumbricus Terrestris Hemoglobin at pH 3.5
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The dissociation of the extracellular hemoglobin from the earthworm Lumbricus terrestris was investigated. Using 8M urea as the dissociation agent, polyacrylamide gel electrophoresis was achieved both in the presence and in the absence of the reducing agent,𝛽-mercaptoethanol. Four bands emerged from the unreduced gels while six bands emerged from the reduced gels. Each of the four unreduced protein bands were removed from the urea gels, reduced in 𝛽-mercaptoethanol and sodium dodecyl sulfate (SDS), and re-electrophoresed through the Laemmli system (SDS was the dissociation agent). The result was that each band coincided very closely with standard hemoglobin bands also reduced in 𝛽-mercaptoethanol and SDS. These results compare favorably to those obtained by Shlom and Vinogradov in 1973 where SDS was the initial denaturant.