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Extra resources for Hemoglobins
8. 52 Aplysia Myoglobin. Myoglobin from the buccal muscles 53 and nerves 54 of the gastropod mollusc Aplysia limacina (Mediterranean) or A. californica (Pacific Coast) lacks a histidine residue distal to the heme. 5r Optical spectra, 58 EPR spectra, 59 ligand affinity,5s'6° kinetics, 61 amino acid sequence, 55 and X-ray diffraction structure 62 are available. Buccal masses [approximately 2 g per animal; 150 ftmol (A. californica) or 500/zmol (A. 0, containing I mM EDTA. The homogenate is centrifuged at 20,000 g for 10 min and fractionated by the addition of solid ammonium sulfate between 50 and 90% saturation, keeping the pH near 8 by the dropwise addition of ammonia.
J. Biochem. 19, 42 (1971). 45 G. Steffens, G. Buse, and A. Wollmer, Eur. J. Biochem. 72, 201 (1977). 4e A. Wollmer, G. St¢ffens, and G. Buse, Eur. J. Biochem. 72, 207 (1977). 4r V. Braun, R. R. Crichton, and G. Braunitzer, Hoppe-Seyler's Z. Physiol. Chem. 349, 197 (1968). 0. 75M phosphate buffer for X-ray diffraction studies. 1 M dithionite in the same buffer, and the carbon monoxide derivative by subsequent exposure to gaseous carbon monoxide. Oxyhemoglobin crystals were obtained by exposing crystals of aquoferric hemoglobin to a mixture of hydrogen sulfide (a reducing agent) and oxygen in the proportion 1:6.
The choice of preparative procedure depends on the use to which the 2 H. Theorell, Biochem. Z. 252, 1 (1932). 3 j. C. Kendrew, Proc. R. Soc. (London), Set. A 201, 62 (1950). 4 j. C. Kendrew and R. G. Parrish, Proc. R. Soc. (London), Ser. A 238, 305 (1956). 5 M. Z. Atassi, Biochim. Biophys. Acta 221, 612 (1970). s A. Akeson and H. Theorell, Arch. Biochem. Biophys. 91, 319 (1960). 7 K. D. Hardman, E. H. Eylar, D. K. Ray, L. J. Banaszak, and F. R. N. Gurd, J. Biol. Chem. 241,432 (1966). s K. D. Hapner, R.