Quaternary structure and functional properties of Penaeus monodon hemocyanin.
The hemocyanin of the tiger shrimp, Penaeus monodon, was investigated with respect to stability and oxygen binding. While hexamers occur as a major component, dodecamers and traces of higher aggregates are also found. Both the hexamers and dodecamers were found to be extremely stable against dissociation at high pH, independently of the presence of calcium ions, in contrast to the known crustacean hemocyanins. This could be caused by only a few additional noncovalent interactions between amino acids located at the subunit-subunit interfaces. Based on X-ray structures and sequence alignments of related hemocyanins, the particular amino acids are identified. At all pH values, the p50 and Bohr coefficients of the hexamers are twice as high as those of dodecamers. While the oxygen binding of hexamers from crustaceans can normally be described by a simple two-state model, an additional conformational state is needed to describe the oxygen-binding behaviour of Penaeus monodon hemocyanin within the pH range of 7.0 to 8.5. The dodecamers bind oxygen according to the nested Monod-Whyman-Changeaux (MWC) model, as observed for the same aggregation states of other hemocyanins. The oxygen-binding properties of both the hexameric and dodecameric hemocyanins guarantee an efficient supply of the animal with oxygen, with respect to the ratio between their concentrations. It seems that under normoxic conditions, hexamers play the major role. Under hypoxic conditions, the hexamers are expected not to be completely loaded with oxygen. Here, the dodecamers are supposed to be responsible for the oxygen supply.
Beltramini
Mariano
M
Department of Biology, University of Padova, Padova, Italy. beltmar@civ.bio.unipd.it
Colangelo
Nadia
N
Giomi
Folco
F
Bubacco
Luigi
L
Di Muro
Paolo
P
Hellmann
Nadja
N
Jaenicke
Elmar
E
Decker
Heinz
H
D000494:Allosteric Regulation; D000818:Animals; D018360:Crystallography, X-Ray; D006433:Hemocyanins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000378:metabolism*; D033561:Penaeidae / Q000737:chemistry*; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D016415:Sequence Alignment
D006433:Hemocyanins; D010100:Oxygen