title: A nonprotein thermal hysteresis-producing xylomannan antifreeze in the freeze-tolerant Alaskan beetle Upis ceramboides.
abstract: Thermal hysteresis (TH), a difference between the melting and freezing points of a solution that is indicative of the presence of large-molecular-mass antifreezes (e.g., antifreeze proteins), has been described in animals, plants, bacteria, and fungi. Although all previously described TH-producing biomolecules are proteins, most thermal hysteresis factors (THFs) have not yet been structurally characterized, and none have been characterized from a freeze-tolerant animal. We isolated a highly active THF from the freeze-tolerant beetle, Upis ceramboides, by means of ice affinity. Amino acid chromatographic analysis, polyacrylamide gel electrophoresis, UV-Vis spectrophotometry, and NMR spectroscopy indicated that the THF contained little or no protein, yet it produced 3.7 +/- 0.3 degrees C of TH at 5 mg/ml, comparable to that of the most active insect antifreeze proteins. Compositional and structural analyses indicated that this antifreeze contains a beta-mannopyranosyl-(1-->4) beta-xylopyranose backbone and a fatty acid component, although the lipid may not be covalently linked to the saccharide. Consistent with the proposed structure, treatment with endo-beta-(1-->4)xylanase ablated TH activity. This xylomannan is the first TH-producing antifreeze isolated from a freeze-tolerant animal and the first in a new class of highly active THFs that contain little or no protein.
authors.affiliation: Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA. kwalter2@nd.edu
authors.forename: Kent R
authors.initials: KR
authors.lastname: Walters
authors.forename: Anthony S
authors.initials: AS
authors.lastname: Serianni
authors.forename: Todd
authors.initials: T
authors.lastname: Sformo
authors.forename: Brian M
authors.initials: BM
authors.lastname: Barnes
authors.forename: John G
authors.initials: JG
authors.lastname: Duman
mesh_terms: D000220:Adaptation, Biological / Q000502:physiology*; D000413:Alaska; D000818:Animals; D021301:Antifreeze Proteins / Q000737:chemistry*; D001517:Coleoptera / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013056:Spectrophotometry, Ultraviolet
chemical_list: D021301:Antifreeze Proteins