title: Spruce budworm antifreeze protein: changes in structure and dynamics at low temperature.
abstract: Antifreeze proteins (AFPs) prevent the growth of ice, and are used by some organisms that live in sub-zero environments for protection against freezing. All AFPs are thought to function by an adsorption inhibition process. In order to elucidate the ice-binding mechanism, the structures of several AFPs have been determined, and have been shown to consist of different folds. Recently, the first structures of the highly active insect AFPs have been characterized. These proteins have a beta-helix structure, which adds yet another fold to the AFP family. The 90-residue spruce budworm (Choristoneura fumiferana) AFP consists of a beta-helix with 15 residues per coil. The structure contains two ranks of aligned threonine residues (known as the TXT motif), which were shown by mutagenesis experiments to be located in the ice-binding face. In our previous NMR study of this AFP at 30 degrees C, we found that the TXT face was not optimally defined because of the broadening of NMR resonances potentially due to weak oligomerization. We present here a structure of spruce budworm AFP determined at 5 degrees C, where this broadening is reduced. In addition, the 1H-15N NMR dynamics of the protein were examined at 30 degrees C and 5 degrees C. The results show that the spruce budworm AFP is more structured at 5 degrees C, and support the general observation that AFPs become more rigid as the temperature is lowered.
authors.affiliation: Department of Biochemistry, CIHR Group in Structure and Function, University of Alberta, 713, Heritage Medical Research Building, T6G 2H7, Edmonton, Alta., Canada.
authors.forename: Steffen P
authors.initials: SP
authors.lastname: Graether
authors.forename: Stéphane M
authors.initials: SM
authors.lastname: Gagné
authors.forename: Leo
authors.initials: L
authors.lastname: Spyracopoulos
authors.forename: Zongchao
authors.initials: Z
authors.lastname: Jia
authors.forename: Peter L
authors.initials: PL
authors.lastname: Davies
authors.forename: Brian D
authors.initials: BD
authors.lastname: Sykes
mesh_terms: D000818:Animals; D021301:Antifreeze Proteins / Q000737:chemistry*; D003080:Cold Temperature; D008958:Models, Molecular; D009036:Moths; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation
chemical_list: D021301:Antifreeze Proteins