A potential role for water in the modulation of oxygen-binding by tarantula hemocyanin.
Hemocyanin from the tarantula Eurypelma californicum is a large respiratory protein with an exceptional high cooperativity. In contrast to hemocyanins from other species, no physiological allosteric effectors other than protons have been identified so far for this 24-meric oligomer. Here we report for the first time the mediating effects of water activity on the oxygen binding properties of a hemocyanin. Oxygen binding curves were measured in presence of several concentrations of glycine and sucrose since both substances reduce water activity. A pronounced shift of the p(50) was observed in both cases but in different directions: adding sucrose shifts the p(50) towards lower values whereas presence of glycine shows the same tendency as for human hemoglobin. Furthermore, prolonged incubation in sucrose slightly distorts the oxygen binding characteristics of spider hemocyanin. Therefore, only the influence of glycine was further analysed. An analysis based on the nested MWC model indicates, that presence of glycine leads to a preferential population of the two states with lower oxygen affinity (tR and tT) compared to the high affinity states rT and rR. The results corroborate the presence of hierarchically organized interactions in this hemocyanin.
Hellmann
N
N
Institute for Molecular Biophysics, University of Mainz, Jakob-Welder-Weg 26, Mainz 55128, Germany. nadja@biophysik.biologie.uni-mainz.de
Raithel
K
K
Decker
H
H
D000818:Animals; D006433:Hemocyanins / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D010100:Oxygen / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D013112:Spiders; D014867:Water / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
D014867:Water; D006433:Hemocyanins; D010100:Oxygen