Abstract
Publisher Summary This chapter focuses on the carbamylation of the amino groups of hemoglobin. The procedure described is only one of three methods that can be employed to prepare gram amounts of hemoglobin carbamylated at specific NH 2 -terminal positions. Hemoglobin obtained from a lysate of fresh erythrocytes in the oxygenated form is the starting material; purification to homogeneity is achieved later by the chromatography of the separate α and β chains after carbamylation. Blood from normal individuals, from patients with sickle cell anemia, or from patients heterozygous for abnormal hemoglobin Providence (Lys- β 82 → Asn or Asp) has been used successfully in the procedure. The chapter describes the use of specifically carbamylated hemoglobin tetramers, removal of minor hemoglobins, preparation and recombination of hemoglobin chains, and conversion of carbon monoxyhemoglobin to oxyhemoglobin. For the isolation of the hemoglobin chains, a modification of the procedure of Bucci and Fronticelli is used.
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