Abstract
In this chapter an overview of the most commonly used techniques in both quantitative and qualitative antifreeze protein (AFP) research is presented. This includes, among others, thermal hysteresis measurement by nanoliter cryoscopy, ice recrystallization inhibition assay by microscopy and immunolocalization. AFP research has come a long way, from merely observing the effects of AFP to explaining some of the fundamental mechanisms of action. These involve a dynamic hydration shell around proteins, the nanometer curvature of ice crystals in the presence of AFP, and the flux of water molecules between ice crystals as they recrystallize. The interest in AFP as an additive to food has always been high, and a wealth of methods have been applied to foods with added AFP. Most of these are not concerned with the function of AFP, but rather the effect on a complex matrix, such as meat or dough. Some of these indirect methods for detecting the effects of AFP are also presented at the end of the chapter.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Baier-Schenk A, Handschin S, Conde-Petit B (2005) Ice in prefermented frozen bread dough - an investigation based on calorimetry and microscopy. Cereal Chem 82(3):251–255. https://doi.org/10.1094/CC-82-0251, wOS:000229284300004
Bar-Dolev M, Celik Y, Wettlaufer JS, Davies PL, Braslavsky I (2012) New insights into ice growth and melting modifications by antifreeze proteins. J R Soc Interface 9(77):3249–3259. https://doi.org/10.1098/rsif.2012.0388
Basu K, Garnham CP, Nishimiya Y, Tsuda S, Braslavsky I, Davies P (2014) Determining the ice-binding planes of antifreeze proteins by fluorescence-based ice plane affinity. J Vis Exp 83:e51185. https://doi.org/10.3791/51185
Basu K, Wasserman SS, Jeronimo PS, Graham LA, Davies PL (2016) Intermediate activity of midge antifreeze protein is due to a tyrosine-rich ice-binding site and atypical ice plane affinity. FEBS J 283(8):1504–1515. https://doi.org/10.1111/febs.13687
Boonsupthip W, Lee TC (2003) Application of antifreeze protein for food preservation: effect of type III antifreeze protein for preservation of gel-forming of frozen and chilled actomyosin. J Food Sci 68(5):1804–1809. https://doi.org/10.1111/j.1365-2621.2003.tb12333.x, wOS:000183947000043
Braslavsky I, Drori R (2013) LabVIEW-operated novel nanoliter osmometer for ice binding protein investigations. J Vis Exp 72:e4189. https://doi.org/10.3791/4189
Buch JL, Ramløv H (2016) An open source cryostage and software analysis method for detection of antifreeze activity. Cryobiology 72(3):251–257. https://doi.org/10.1016/j.cryobiol.2016.03.010
Buch J, Ramløv H (2017) Detecting seasonal variation of antifreeze protein distribution in Rhagium mordax using immunofluorescence and high resolution microscopy. Cryobiology 74:132–140. https://doi.org/10.1016/j.cryobiol.2016.11.003
Budke C, Heggemann C, Koch M, Sewald N, Koop T (2009) Ice recrystallization kinetics in the presence of synthetic antifreeze glycoprotein analogues using the framework of LSW theory. J Phys Chem B 113(9):2865–2873. https://doi.org/10.1021/jp805726e
Budke C, Dreyer A, Jaeger J, Gimpel K, Berkemeier T, Bonin AS, Nagel L, Plattner C, DeVries AL, Sewald N, Koop T (2014) Quantitative efficacy classification of ice recrystallization inhibition agents. Cryst Growth Des 14(9):4285–4294. https://doi.org/10.1021/cg5003308
Bulavin LA, Lokotosh TV, Malomuzh NP (2008) Role of the collective self-diffusion in water and other liquids. J Mol Liq 137(1):1–24. https://doi.org/10.1016/j.molliq.2007.05.003
Cao H, Zhao Y, Zhu YB, Xu F, Yu JS, Yuan M (2016) Antifreeze and cryoprotective activities of ice-binding collagen peptides from pig skin. Food Chem 194:1245–1253. https://doi.org/10.1016/j.foodchem.2015.08.102
Chakrabartty A, Hew CL (1991) The effect of enhanced alpha-helicity on the activity of a winter flounder antifreeze polypeptide. Eur J Biochem 202(3):1057–1063. https://doi.org/10.1111/j.1432-1033.1991.tb16470.x
Ding X, Zhang H, Liu W, Wang L, Qian H, Qi X (2014) Extraction of carrot (Daucus carota) antifreeze proteins and evaluation of their effects on frozen white salted noodles. Food Bioprocess Technol 7(3):842–852. https://doi.org/10.1007/s11947-013-1101-0
Duman JG (1980) Factors involved in overwintering survival of the freeze tolerant beetle, Dendroides canadensis. J Comp Physiol B 136(1):52–59. https://doi.org/10.1007/BF00688622
Easton CM, Horwath KL (1994) Characterization of primary cell cultures derived from fat body of the beetle, Tenebrio molitor, and the immunolocalization of a thermal hysteresis protein in vitro. J Insect Physiol 40(6):537–547. https://doi.org/10.1016/0022-1910(94)90127-9
Ebbinghaus S, Kim SJ, Heyden M, Yu X, Heugen U, Gruebele M, Leitner DM, Havenith M (2007) An extended dynamical hydration shell around proteins. PNAS 104(52):20749–20752. https://doi.org/10.1073/pnas.0709207104
Ebbinghaus S, Meister K, Born B, DeVries AL, Gruebele M, Havenith M (2010) Antifreeze glycoprotein activity correlates with long-range protein-water dynamics. J Am Chem Soc 132(35):12210–12211. https://doi.org/10.1021/ja1051632
Ebbinghaus S, Meister K, Prigozhin MB, DeVries AL, Havenith M, Dzubiella J, Grue-bele M (2012) Functional importance of short-range binding and long-range solvent interactions in helical antifreeze peptides. Biophys J 103(2):L20–L22. https://doi.org/10.1016/j.bpj.2012.06.013
Esser-Kahn AP, Trang V, Francis MB (2010) Incorporation of antifreeze proteins into polymer coatings using site-selective bioconjugation. J Am Chem Soc 132(38):13264–13269. https://doi.org/10.1021/ja103038p
Evans RP, Hobbs RS, Goddard SV, Fletcher GL (2007) The importance of dissolved salts to the in vivo efficacy of antifreeze proteins. Comp Biochem Physiol A 148(3):556–561. https://doi.org/10.1016/j.cbpa.2007.07.005
Gaede-Koehler A, Kreider A, Canfield P, Kleemeier M, Grunwald I (2012) Direct measurement of the thermal hysteresis of antifreeze proteins (AFPs) using sonocrystallization. Anal Chem 84(23):10229–10235. https://doi.org/10.1021/ac301946w, wOS:000311815300014
Garnham CP, Natarajan A, Middleton AJ, Kuiper MJ, Braslavsky I, Davies PL (2010) Compound ice-binding site of an antifreeze protein revealed by mutagenesis and fluorescent tagging. Biochemistry 49(42):9063–9071. https://doi.org/10.1021/bi100516e
Graether SP, Kuiper MJ, Gagne SM, Walker VK, Jia Z, Sykes BD, Davies PL (2000) Beta-helix structure and ice-binding properties of a hyperactive antifreeze protein from an insect. Nature 406(6793):325–328
Grandum S, Yabe A, Nakagomi K, Tanaka M, Takemura F, Kobayashi Y, Frivik PE (1999) Analysis of ice crystal growth for a crystal surface containing adsorbed antifreeze proteins. J Cryst Growth 205(3):382–390. https://doi.org/10.1016/S0022-0248(99)00267-5, wOS:000082217200018
Gwak Y, Ji P, Kim M, Kim HS, Kwon MJ, Oh SJ, Kim YP, Jin E (2015) Creating anti-icing surfaces via the direct immobilization of antifreeze proteins on aluminum. Sci Rep 5:12019. https://doi.org/10.1038/srep12019
Halwani DO, Brockbank KGM, Duman JG, Campbell LH (2014) Recombinant Dendroides canadensis antifreeze proteins as potential ingredients in cryopreservation solutions. Cryobiology 68(3):411–418. https://doi.org/10.1016/j.cryobiol.2014.03.006
Hansen TN, Baust JG (1988) Differential scanning calorimetric analysis of antifreeze protein activity in the common mealworm, Tenebrio molitor. BBA-Protein Struct M 957(2):217–221. https://doi.org/10.1016/0167-4838(88)90275-0
Hassa-Roudsari M, Goff HD (2012) A new quantitative method to measure activity of ice structuring proteins using differential scanning calorimetry. Cryoletters 33(2):118–125
Haymet ADJ, Ward LG, Harding MM, Knight CA (1998) Valine substituted winter flounder ‘antifreeze’: preservation of ice growth hysteresis. FEBS Lett 430(3):301–306. https://doi.org/10.1016/S0014-5793(98)00652-8, wOS:000074797600034
Haymet ADJ, Ward LG, Harding MM (1999) Winter flounder “antifreeze” proteins: synthesis and ice growth inhibition of analogues that probe the relative importance of hydrophobic and hydrogen-bonding interactions. J Am Chem Soc 121(5):941–948. https://doi.org/10.1021/ja9801341
Heisig M, Mattessich S, Rembisz A, Acar A, Shapiro M, Booth CJ, Neelakanta G, Fikrig E (2015) Frostbite protection in mice expressing an antifreeze glycoprotein. PLoS One 10(2):e0116562. https://doi.org/10.1371/journal.pone.0116562
Hon WC, Griffith M, Chong P, Yang DS (1994) Extraction and isolation of antifreeze proteins from winter rye (Secale cereale L.) leaves. Plant Physiol 104(3):971–980
Horwath KL, Easton CM, Poggioli GJ, Myers K, Schnorr IL (1996) Tracking the profile of a specific antifreeze protein and its contribution to the thermal hysteresis activity in cold hardy insects. Eur J Entomol 93(3):419–433, wOS:A1996VN62700013
Ickes L, Welti A, Hoose C, Lohmann U (2015) Classical nucleation theory of homogeneous freezing of water: thermodynamic and kinetic parameters. Phys Chem Chem Phys 17(8):5514–5537. https://doi.org/10.1039/C4CP04184D
Jackman J, Noestheden M, Moffat D, Pezacki JP, Findlay S, Ben RN (2007) Assessing antifreeze activity of AFGP 8 using domain recognition software. Biochem Biophys Res Commun 354(2):340–344. https://doi.org/10.1016/j.bbrc.2006.12.225
Kim EJ, Lee JH, Lee SG, Han SJ (2017) Improving thermal hysteresis activity of antifreeze protein from recombinant Pichia pastoris by removal of N-glycosylation. Prep Biochem Biotechnol 47(3):299–304. https://doi.org/10.1080/10826068.2016.1244682
Knight CA (1966) Formation of crystallographic etch pits on ice, and its application to the study of hailstones. J Appl Meteorol 5(5):710–714
Knight CA, Hallett J, DeVries AL (1988) Solute effects on ice recrystallization: an assessment technique. Cryobiology 25(1):55–60. https://doi.org/10.1016/0011-2240(88)90020-X
Knight C, Cheng C, DeVries A (1991) Adsorption of alpha-helical antifreeze peptides on specific ice crystal surface planes. Biophys J 59(2):409–418. https://doi.org/10.1016/S0006-3495(91)82234-2
Knight C, Driggers E, DeVries A (1993) Adsorption to ice of fish antifreeze glycopeptides 7 and 8. Biophys J 64(1):252–259. https://doi.org/10.1016/S0006-3495(93)81361-4
Kondo H, Hanada Y, Sugimoto H, Hoshino T, Garnham CP, Davies PL, Tsuda S (2012) Ice-binding site of snow mold fungus antifreeze protein deviates from structural regularity and high conservation. PNAS 109(24):9360–9365. https://doi.org/10.1073/pnas.1121607109
Kontogiorgos V, Goff HD, Kasapis S (2008) Effect of aging and ice-structuring proteins on the physical properties of frozen flour–water mixtures. Food Hydrocoll 22(6):1135–1147. https://doi.org/10.1016/j.foodhyd.2007.06.005
Kristiansen E, Zachariassen KE (2005) The mechanism by which fish antifreeze proteins cause thermal hysteresis. Cryobiology 51(3):262–280. https://doi.org/10.1016/j.cryobiol.2005.07.007
Kristiansen E, Pedersen SA, Zachariassen KE (2008) Salt-induced enhancement of antifreeze protein activity: a salting-out effect. Cryobiology 57(2):122–129. https://doi.org/10.1016/j.cryobiol.2008.07.001
Leygonie C, Britz TJ, Hoffman LC (2012) Impact of freezing and thawing on the quality of meat: review. Meat Sci 91(2):93–98. https://doi.org/10.1016/j.meatsci.2012.01.013
Li N, Andorfer CA, Duman JG (1998) Enhancement of insect antifreeze protein activity by solutes of low molecular mass. J Exp Biol 201(15):2243–2251
Liu J, Xu X, Xu Q, Wang S, Xu J (2014) Transgenic tobacco plants expressing PicW. Plant Cell Tiss Org Cult 118(3):391–400. https://doi.org/10.1007/s11240-014-0491-7
Lv J, Song Y, Jiang L, Wang J (2014) Bio-inspired strategies for anti-icing. ACS Nano 8(4):3152–3169. https://doi.org/10.1021/nn406522n
Mangiagalli M, Bar-Dolev M, Tedesco P, Natalello A, Kaleda A, Brocca S, de Pascale D, Pucciarelli S, Miceli C, Braslavsky I, Lotti M (2017) Cryo-protective effect of an ice-binding protein derived from Antarctic bacteria. FEBS J 284(1):163–177. https://doi.org/10.1111/febs.13965
Marshall CB, Daley ME, Graham LA, Sykes BD, Davies PL (2002) Identification of the icebinding face of antifreeze protein from Tenebrio molitor. FEBS Lett 529(2–3):261–267. https://doi.org/10.1016/S0014-5793(02)03355-0
Meister K, Ebbinghaus S, Xu Y, Duman JG, DeVries A, Gruebele M, Leitner DM, Havenith M (2013) Long-range protein–water dynamics in hyperactive insect antifreeze proteins. PNAS 110(5):1617–1622. https://doi.org/10.1073/pnas.1214911110
Meister K, Duman JG, Xu Y, DeVries AL, Leitner DM, Havenith M (2014a) The role of sulfates on antifreeze protein activity. J Phys Chem B 118(28):7920–7924. https://doi.org/10.1021/jp5006742
Meister K, Strazdaite S, DeVries AL, Lotze S, Olijve LLC, Voets IK, Bakker HJ (2014b) Observation of ice-like water layers at an aqueous protein surface. PNAS 111(50):17732–17736
Mitchell DE, Congdon T, Rodger A, Gibson MI (2015) Gold nanoparticle aggregation as a probe of antifreeze (glyco) protein-inspired ice recrystallization inhibition and identification of new IRI active macromolecules. Sci Rep 5:15716. https://doi.org/10.1038/srep15716
Mizrahy O, Bar-Dolev M, Guy S, Braslavsky I (2013) Inhibition of ice growth and recrystallization by zirconium acetate and zirconium acetate hydroxide. PLoS One 8(3):e59540. https://doi.org/10.1371/journal.pone.0059540
Namperumal R, Coger R (1998) A new cryostage design for cryomicroscopy. J Microsc (Oxford) 192(2):202–211. https://doi.org/10.1046/j.1365-2818.1998.00416.x
Olijve LLC, Meister K, DeVries AL, Duman JG, Guo S, Bakker HJ, Voets IK (2016) Blocking rapid ice crystal growth through nonbasal plane adsorption of antifreeze proteins. PNAS 113(14):3740–3745. https://doi.org/10.1073/pnas.1524109113
Olsen TM, Sass SJ, Li N, Duman JG (1998) Factors contributing to seasonal increases in inoculative freezing resistance in overwintering fire-colored beetle larvae Dendroides canadensis (Pyrochroidae). J Exp Biol 201(10):1585–1594, wOS:000074211300007
Panadero J, Randez-Gil F, Prieto JA (2005) Heterologous expression of type I antifreeze peptide GS-5 in Baker’s yeast increases freeze tolerance and provides enhanced gas production in frozen dough. J Agric Food Chem 53(26):9966–9970. https://doi.org/10.1021/jf0515577
Park JI, Lee JH, Gwak Y, Kim HJ, Jin E, Kim YP (2012a) Frozen assembly of gold nanoparticles for rapid analysis of antifreeze protein activity. Biosens Bioelectron 41:752–757. https://doi.org/10.1016/j.bios.2012.09.052
Park KS, Do H, Lee JH, Park SI, Jung Kim E, Kim SJ, Kang SH, Kim HJ (2012b) Characterization of the ice-binding protein from Arctic yeast Leucosporidium sp. AY30. Cryobiology 64(3):286–296. https://doi.org/10.1016/j.cryobiol.2012.02.014
Payne SR, Young OA (1995) Effects of pre-slaughter administration of antifreeze proteins on frozen meat quality. Meat Sci 41(2):147–155. https://doi.org/10.1016/0309-1740(94)00073-G
Payne SR, Sandford D, Harris A, Young OA (1994) The effects of antifreeze proteins on chilled and frozen meat. Meat Sci 37(3):429–438. https://doi.org/10.1016/0309-1740(94)90058-2
Pertaya N, Celik Y, DiPrinzio CL, Wettlaufer JS, Davies PL, Braslavsky I (2007) Growth–melt asymmetry in ice crystals under the influence of spruce budworm antifreeze protein. J Phys Condens Matter 19(41):412101. https://doi.org/10.1088/0953–8984/19/41/412101
Pertaya N, Marshall CB, Celik Y, Davies PL, Braslavsky I (2008) Direct visualization of spruce budworm antifreeze protein interacting with ice crystals: basal plane affinity confers hyperactivity. Biophys J 95(1):333–341. https://doi.org/10.1529/biophysj.107.125328, wOS:000256668200033
Qiu L, Wang Y, Wang J, Zhang F, Ma J (2009) Expression of biologically active recombinant antifreeze protein His-MpAFP149 from the desert beetle Microdera punctipennis dzungarica in Escherichia coli. Mol Biol Rep 37(4):1725–1732. https://doi.org/10.1007/s11033-009-9594-3
Ramløv H, Wharton DA, Wilson PW (1996) Recrystallization in a freezing tolerant Antarctic nematode, Panagrolaimus davidi, and an Alpine Weta, Hemideina maori (Orthoptera; Stenopelmatidae). Cryobiology 33(6):607–613. https://doi.org/10.1006/cryo.1996.0064
Ramløv H, DeVries A, Wilson P (2005) Antifreeze glycoproteins from the Antarctic fish Dissostichus mawsoni studied by differential scanning calorimetry (DSC) in combination with nanolitre osmometry. Cryoletters 26(2):73–84
Raymond JA, DeVries AL (1977) Adsorption inhibition as a mechanism of freezing resistance in polar fishes. PNAS 74(6):2589–2593
Raymond JA, Wilson P, DeVries AL (1989) Inhibition of growth of nonbasal planes in ice by fish antifreezes. PNAS 86(3):881–885
Romero I, Fernandez-Caballero C, Goñi O, Escribano MI, Merodio C, Sanchez-Ballesta MT (2008) Functionality of a class I beta-1,3-glucanase from skin of table grapes berries. Plant Sci 174(6):641–648. https://doi.org/10.1016/j.plantsci.2008.03.019
Scotter AJ, Marshall CB, Graham LA, Gilbert JA, Garnham CP, Davies PL (2006) The basis for hyperactivity of antifreeze proteins. Cryobiology 53(2):229–239. https://doi.org/10.1016/j.cryobiol.2006.06.006, wOS:000241145300008
Takamichi M, Nishimiya Y, Miura A, Tsuda S (2009) Fully active QAE isoform confers thermal hysteresis activity on a defective SP isoform of type III antifreeze protein. FEBS J 276(5):1471–1479. https://doi.org/10.1111/j.1742-4658.2009.06887.x, wOS:000263451600027
Tomczak MM, Marshall CB, Gilbert JA, Davies PL (2003) A facile method for determining ice recrystallization inhibition by antifreeze proteins. Biochem Biophys Res Commun 311(4):1041–1046. https://doi.org/10.1016/j.bbrc.2003.10.106
Tong L, Lin Q, Wong WKR, Ali A, Lim D, Sung WL, Hew CL, Yang DSC (2000) Extracellular expression, purification, and characterization of a winter flounder antifreeze polypeptide from Escherichia coli. Protein Expr Purif 18(2):175–181. https://doi.org/10.1006/prep.1999.1176
Wang S, Amornwittawat N, Banatlao J, Chung M, Kao Y, Wen X (2009a) Hofmeister effects of common monovalent salts on the beetle antifreeze protein activity. J Phys Chem B 113(42):13891–13894. https://doi.org/10.1021/jp907762u, wOS:000270670800029
Wang S, Amornwittawat N, Juwita V, Kao Y, Duman JG, Pascal TA, Goddard WA, Wen X (2009b) Arginine, a key residue for the enhancing ability of an antifreeze protein of the beetle Dendroides canadensis. Biochemistry 48(40):9696–9703. https://doi.org/10.1021/bi901283p
Wen D, Laursen R (1992) A model for binding of an antifreeze polypeptide to ice. Biophys J 63(6):1659–1662, wOS:A1992KF55100025
Wilson P, Gould M, DeVries A (2002) Hexagonal shaped ice spicules in frozen antifreeze protein solutions. Cryobiology 44(3):240–250. https://doi.org/10.1016/S0011-2240(02)00028-7
Wu DW, Duman JG (1991) Activation of antifreeze proteins from larvae of the beetle Dendroides canadensis. J Comp Physiol B 161(3):279–283. https://doi.org/10.1007/BF00262309
Xu Y, Bäumer A, Meister K, Bischak CG, DeVries AL, Leitner DM, Havenith M (2016) Protein-water dynamics in antifreeze protein III activity. Chem Phys Lett 647:1–6. https://doi.org/10.1016/j.cplett.2015.11.030
Yeh CM, Kao BY, Peng HJ (2009) Production of a recombinant type 1 antifreeze protein analogue by L. lactis and its applications on frozen meat and frozen dough. J Agric Food Chem 57(14):6216–6223. https://doi.org/10.1021/jf900924f
Zhang C, Zhang H, Wang L (2007a) Effect of carrot (Daucus carota) antifreeze proteins on the fermentation capacity of frozen dough. Food Res Int 40(6):763–769. https://doi.org/10.1016/j.foodres.2007.01.006, wOS:000247326000014
Zhang C, Zhang H, Wang L, Gao H, Guo XN, Yao HY (2007b) Improvement of texture properties and flavor of frozen dough by carrot (Daucus carota) antifreeze protein supplementation. J Agric Food Chem 55(23):9620–9626. https://doi.org/10.1021/jf0717034
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Nature Switzerland AG
About this chapter
Cite this chapter
Buch, J.L. (2020). Measuring Antifreeze Protein Activity. In: Ramløv, H., Friis, D. (eds) Antifreeze Proteins Volume 2. Springer, Cham. https://doi.org/10.1007/978-3-030-41948-6_9
Download citation
DOI: https://doi.org/10.1007/978-3-030-41948-6_9
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-030-41947-9
Online ISBN: 978-3-030-41948-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)