Purification, crystallisation and preliminary X-ray study of haemoglobin from Crocodilis palustris and Crocodilis porosus

摘要

The unsolved three-dimensional structure of crocodile haemoglobin and its prospects as a blood substitute have led us to initiate the purification and crystallisation of haemoglobin molecules from crocodile species (Crocodilis palustris or mugger and Crocodilis porosus or salt water crocodile). The work has resulted in the prevention of polymerisation of naked haemoglobin molecules using N-ethylmaleimide or iodoacetamide. The purified monomeric haemoglobin molecule of C. porosus was crystallised in two different forms and X-ray diffraction data were collected up to 2 A resolution for both forms. Form I: a = 53.62, b = 53.55, c = 103.77 A; β= 93.35°, space group P2_1, Z = 2. Form II: a = 71.30, b = 54.70, c = 80.00 A; β= 106.4°, space group P2_1, Z = 2. Structure solution and rigid body refinement of form I data resulted in a model with R_(free) = 0.42 and R = 0.35.