Crystallization and diffraction patterns of the oxy and cyano forms of the Lucina pectinata haemoglobins complex.
The native oxygen-carrier haemoglobins complex (HbII-III) is composed of haemoglobin II (HbII) and haemoglobin III (HbIII), which are found in the ctenidia tissue of the bivalve mollusc Lucina pectinata. This protein complex was isolated and purified from its natural source and crystallized using the vapour-diffusion and capillary counter-diffusion methods. Oxy and cyano derivatives of the complex crystallized using several conditions, but the best crystals in terms of quality and size were obtained from sodium formate pH 5 using the counter-diffusion method in a single capillary. Crystals of the oxy and cyano complexes, which showed a ruby-red colour and nonsingular prismatic shapes, scattered X-rays to resolution limits of 2.15 and 2.20 A, respectively, using a 0.886 A synchrotron-radiation source. The crystals belonged to the tetragonal system, space group P4(2)2(1)2, with unit-cell parameters a = b = 74.07, c = 152.07 and a = b = 73.83, c = 152.49 A for the oxy and cyano complexes, respectively. The asymmetric unit of both crystals is composed of a single copy of the heterodimer, with Matthew coefficients (V(M)) of 3.08 and 3.06 A(3) Da(-1) for the oxy and cyano complexes, respectively, which correspond to a solvent content of approximately 60.0% by volume.
Ruiz-Martínez
Carlos R
CR
Departamento de Química, Universidad de Puerto Rico, Recinto de Mayagüez.
Nieves-Marrero
Carlos A
CA
Estremera-Andújar
Rafael A
RA
Gavira
José A
JA
González-Ramírez
Luis A
LA
López-Garriga
Juan
J
García-Ruiz
Juan M
JM
D000818:Animals; D018360:Crystallography, X-Ray / Q000379:methods*; D004058:Diffusion; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008024:Ligands; D015233:Models, Statistical; D008974:Mollusca; D010100:Oxygen / Q000737:chemistry; D011487:Protein Conformation; D014961:X-Ray Diffraction / Q000379:methods*
D006454:Hemoglobins; D008024:Ligands; D006418:Heme; D010100:Oxygen