ORIGINAL PAPER A thermal hysteresis-producing xylomannan glycolipid antifreeze associated with cold tolerance is found in diverse taxa Kent R. Walters Jr. • Anthony S.Serianni • Yann Voituron • Todd Sformo • Brian M. Barnes • John G. Duman Received: 12 November 2010 / Revised: 28 December 2010 / Accepted: 5 January 2011 / Published online: 30 January 2011 Ó Springer-Verlag 2011 Abstract The presence of large-molecular-mass, thermal hysteresis (TH)-producing antifreezes (e.g., antifreeze pro- teins)hasbeen reported in numerous and diverse taxa, including representative species of fish, arthropods, plants, fungi,and bacteria. However, relatively few of these anti- freeze molecules have been chemically characterized. We screened diverse species by subjecting their homogenates to ice-affinity purification and discovered the presence of a newly identified class of antifreeze, a xylomannan-based TH-producing glycolipid that was previously reported in one species of freeze-tolerant Alaskan beetle. We isolated xylomannan-based antifreeze glycolipids from one plant species, six insect species, and the first frog species to be shown to produce a large-molecular-mass antifreeze. 1 H NMR spectra of the ice-purified molecules isolated from these diverse freeze-tolerant and freeze-avoiding organism were nearly identical, indicating that the chemical structu of the glycolipids were highly similar. Although the exact functions remain uncertain, it appears that antifreeze gly- colipids play a role in cold tolerance. Keywords Freeze tolerance Antifreeze proteins Cryoprotection Cold adaptation Antifreeze glycolipid Abbreviations TH Thermal hysteresis AF(G)Ps Antifreeze (glyco)proteins AFGLs Antifreeze glycolipids Introduction Subzero winter temperatures pose a significant challenge the survival of organisms in temperate and polar regions. response, many poikilothermic organisms living in these areas,including fishes, amphibians, reptiles,arthropods, plants,fungi and bacteria, haveevolved physiological adaptations to survive subzero temperatures (Denlinger a Lee 2010;DeVries and Cheng 2005; Duman and Olsen 1993;Robinson 2001; Storey 1990; Thomashow 1999; Voituron etal. 2005).In a given organism, multiple physiological adaptations permit cold hardiness, including Communicated by H.V. Carey. K. R. Walters Jr.J. G. Duman Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA A. S. Serianni Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA Y. Voituron Physiologie des re ´gulations e ´nerge ´tiques, cellulaires, et mole ´culaires (UMR CNRS 5123), Universite ´ Claude Bernard Lyon1, Villeurbanne, France T. Sformo B.M. Barnes Institute of Arctic Biology, University of Alaska Fairbanks, Fairbanks, AK 99775,USA Present Address: K. R. Walters Jr. (&) Department of Entomology, University of Illinois at Urbana-Champaign, 320 Morrill Hall, 505 S Goodwin Avenue, Urbana, IL 61801,USA e-mail: kwalte2@life.illinois.edu Present Address: T. Sformo Department of Wildlife Management, North Slope Borough, Barrow,AK 99723,USA 123 J Comp Physiol B (2011) 181:631–640 DOI 10.1007/s00360-011-0552-8