Naturally crystalline hemoglobin of the nematode Mermis nigrescens. An in situ microspectrophotometric study of chemical properties and dichroism.
A dichroic microspectrophotometer was used to measure isotropic and dichroic absorbance spectra of this unique cytoplasmic hemoglobin and its derivatives. A perfusion slide enabled changing the media bathing the Mermis head. The native spectrum, which has an exceptionally low alpha-band extinction, was shown to be entirely due to oxyhemoglobin. The CO-hemoglobin spectrum is more typical, however, the alpha- and beta-bands are unusually closely spaced. A ferric hemochrome was formed on oxidation with ferricyanide or hydroxylamine and was readily converted to ferric hemoglobin cyanide on adding cyanide. Aquoferric hemoglobin and ferric hemoglobin fluoride were not easily formed. Deoxyhemoglobin, identified by its typical absorption spectrum, was formed only under the extremely low O2 pressures attainable in the presence of dithionite. A glucose oxidase, catalase solution deoxygenated hemoglobin in human erythrocytes but not in adjacent Mermis preparations. The affinity for O2 is much greater than for CO. Also, spectral evidence points to an oxyheme environment that is different than in vertebrate hemoglobin and myoglobin. The polarization ratio (PR) magnitude and the PR spectrum were unaffected by perfusion with high refractive index solvents; therefore, form dichroism due to the rodlike crystals is negligible. Maximum extinction is approximately perpendicular to the long axis of the microscopic crystals, which are oriented parallel to the body axis within the hypodermal cells. The PR spectra of the hemoglobin derivatives strongly resemble the corresponding spectra previously reported of single crystals made of horse hemoglobin, whale myoglobin, or Aplysia myoglobin and change appropriately when the ligand is changed. This confirms that the intracellular crystals of Mermis are of oxyhemoglobin.
D000818:Animals; D002248:Carbon Monoxide / Q000378:metabolism; D002263:Carboxyhemoglobin; D003460:Crystallization; D004227:Dithionite / Q000494:pharmacology; D005260:Female; D006454:Hemoglobins / Q000378:metabolism*; D008706:Methemoglobin / Q000031:analogs & derivatives; D009348:Nematoda / Q000032:analysis*; D010100:Oxygen / Q000378:metabolism; D010108:Oxyhemoglobins; D013057:Spectrum Analysis
D006454:Hemoglobins; D010108:Oxyhemoglobins; C007169:cyanomethemoglobin; D004227:Dithionite; D002248:Carbon Monoxide; C100003:deoxyhemoglobin; D008706:Methemoglobin; D002263:Carboxyhemoglobin; D010100:Oxygen