Thermal stability of homologous functional units of Helix pomatia hemocyanin does not correlate with carbohydrate content.
The thermal stability of the eight functional units of beta-hemocyanin of the gastropodan mollusc Helix pomatia was investigated by FTIR spectroscopy. Molluscan hemocyanin functional units have a molecular mass of approximately 50 kDa and generally contain three disulfide bridges: two in the mainly alpha-helical N-terminal domain and one in the C-terminal beta-sheet domain. They show more than 50% sequence homology and it is assumed that they adopt a similar conformation. However, the functional units of H. pomatiabeta-hemocyanin, designated HpH-a to HpH-h, differ considerably in their carbohydrate content (0-18 wt%). Most functional units are exceptionally stable with a melting temperature in the range 77-83 degrees C. Two functional units, HpH-b and HpH-c, however, have a reduced stability with melting temperature values of 73 degrees C and 64 degrees C, respectively. Although the most glycosylated functional unit (HpH-g) has the highest temperature stability, there is no linear correlation between the degree of glycosylation of the functional units and the unfolding temperature. This is ascribed to variations in secondary structure as well as in glycan attachment sites. Moreover, the disulfide bonds might play an important role in the conformational stability of the functional units. Sequence comparison of molluscan hemocyanins suggests that the less stable functional units, HpH-b and HpH-c, similar to most of their paralogous counterparts, lack the disulfide bond in the C-terminal domain.
Division of Biochemistry, Molecular and Structural Biology, Department of Chemistry, Katholieke Universiteit Leuven, Belgium.
Betül T
BT
Yesilyurt
Constant
C
Gielens
Filip
F
Meersman
D000818:Animals; D002241:Carbohydrates / Q000032:analysis*; D006372:Helix, Snails; D006433:Hemocyanins / Q000737:chemistry*; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature
D002241:Carbohydrates; D021122:Protein Subunits; D006433:Hemocyanins