title: The importance of size and disorder in the cryoprotective effects of dehydrins.
abstract: Dehydrins protect plant proteins and membranes from damage during drought and cold. Vitis riparia K2 is a 48-residue protein that can protect lactate dehydrogenase from freeze-thaw damage by preventing the aggregation and denaturation of the enzyme. To further elucidate its mechanism, we used a series of V. riparia K2 concatemers (K4, K6, K8, and K10) and natural dehydrins (V. riparia YSK2, 60 kilodalton peach dehydrin [PCA60], barley dehydrin5 [Dhn5], Thellungiella salsuginea dehydrin2 [TsDHN-2], and Opuntia streptacantha dehydrin1 [OpsDHN-1]) to test the effect of the number of K-segments and dehydrin size on their ability to protect lactate dehydrogenase from freeze-thaw damage. The results show that the larger the hydrodynamic radius of the dehydrin, the more effective the cryoprotection. A similar trend is observed with polyethylene glycol, which would suggest that the protection is simply a nonspecific volume exclusion effect that can be manifested by any protein. However, structured proteins of a similar range of sizes did not show the same pattern and level of cryoprotection. Our results suggest that with respect to enzyme protection, dehydrins function primarily as molecular shields and that their intrinsic disorder is required for them to be an effective cryoprotectant. Lastly, we show that the cryoprotection by a dehydrin is not due to any antifreeze protein-like activity, as has been reported previously.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Hughes
authors.forename: Verena
authors.initials: V
authors.lastname: Schart
authors.forename: Janet
authors.initials: J
authors.lastname: Malcolmson
authors.forename: Kaley A
authors.initials: KA
authors.lastname: Hogarth
authors.forename: David M
authors.initials: DM
authors.lastname: Martynowicz
authors.forename: Erik
authors.initials: E
authors.lastname: Tralman-Baker
authors.forename: Shruti N
authors.initials: SN
authors.lastname: Patel
authors.forename: Steffen P
authors.initials: SP
authors.lastname: Graether
mesh_terms: D021301:Antifreeze Proteins / Q000737:chemistry / Q000494:pharmacology; D055162:Biocatalysis / Q000187:drug effects; D002942:Circular Dichroism; D003451:Cryoprotective Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D003460:Crystallization; D005615:Freezing; D001467:Hordeum / Q000235:genetics / Q000378:metabolism; D007053:Ice; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D027861:Prunus / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology*; D027843:Vitis / Q000235:genetics / Q000378:metabolism
chemical_list: D021301:Antifreeze Proteins; D003451:Cryoprotective Agents; D007053:Ice; D010455:Peptides; D010940:Plant Proteins; D011994:Recombinant Proteins; C070556:dehydrin proteins, plant; D007770:L-Lactate Dehydrogenase
references.citation: Biochem Cell Biol. 2010 Apr;88(2):223-9
references.pmid: 20453925
references.citation: Biochemistry. 1967 Nov;6(11):3489-97
references.pmid: 6073034
references.citation: Biophys J. 2009 Jul 22;97(2):590-8
references.pmid: 19619474
references.citation: Biochemistry (Mosc). 2003 Sep;68(9):945-51
references.pmid: 14606934
references.citation: Trends Biochem Sci. 2002 Feb;27(2):101-6
references.pmid: 11852248
references.citation: Eur J Biochem. 2002 Jan;269(1):2-12
references.pmid: 11784292
references.citation: Plant Physiol. 2006 Jun;141(2):638-50
references.pmid: 16565295
references.citation: Biol Chem. 1996 Sep;377(9):555-61
references.pmid: 9067253
references.citation: Biophys J. 2006 Sep 15;91(6):2243-9
references.pmid: 16798808
references.citation: Plant Cell Environ. 2008 Dec;31(12):1781-90
references.pmid: 18761701
references.citation: Eur J Biochem. 2004 Aug;271(16):3285-96
references.pmid: 15291806
references.citation: Protein Sci. 2002 Apr;11(4):739-56
references.pmid: 11910019
references.citation: Anal Biochem. 1987 Apr;162(1):47-64
references.pmid: 3605596
references.citation: Biochem J. 2005 May 15;388(Pt 1):151-7
references.pmid: 15631617
references.citation: Plant Cell Environ. 2009 May;32(5):532-41
references.pmid: 19183287
references.citation: Planta. 2012 Mar;235(3):565-78
references.pmid: 21984262
references.citation: Cryobiology. 1995 Feb;32(1):23-34
references.pmid: 7697996
references.citation: J Mol Biol. 2003 Apr 11;327(5):1155-68
references.pmid: 12662938
references.citation: Biochim Biophys Acta. 2012 Jul;1824(7):891-7
references.pmid: 22579671
references.citation: Biochem J. 1992 Mar 1;282 ( Pt 2):589-93
references.pmid: 1546973
references.citation: Cell Mol Biol Lett. 2006;11(4):536-56
references.pmid: 16983453
references.citation: J Plant Physiol. 2012 Oct 15;169(15):1559-64
references.pmid: 22902206
references.citation: Biochem Cell Biol. 2010 Oct;88(5):791-807
references.pmid: 20921991
references.citation: Physiol Plant. 2008 Dec;134(4):583-97
references.pmid: 19000195
references.citation: Plant J. 1995 Oct;8(4):583-93
references.pmid: 7496403
references.citation: Plant Cell. 1994 Mar;6(3):351-60
references.pmid: 8180497
references.citation: Methods Biochem Anal. 1970;18:1-53
references.pmid: 4909316
references.citation: Arch Biochem Biophys. 1996 Aug 15;332(2):231-8
references.pmid: 8806730
references.citation: Biosci Biotechnol Biochem. 2000 Aug;64(8):1656-63
references.pmid: 10993152
references.citation: Plant Physiol. 2009 Jul;150(3):1503-14
references.pmid: 19439573
references.citation: J Dairy Sci. 2002 Nov;85(11):2711-21
references.pmid: 12487438
references.citation: Mol Biosyst. 2012 Jan;8(1):210-9
references.pmid: 21909508
references.citation: J Biol Chem. 2003 Oct 17;278(42):40882-9
references.pmid: 12917402
references.citation: Biomol NMR Assign. 2009 Dec;3(2):273-5
references.pmid: 19842064
references.citation: Plant Cell Rep. 2006 Sep;25(9):968-77
references.pmid: 16552595
references.citation: Biosci Biotechnol Biochem. 2003 Aug;67(8):1832-5
references.pmid: 12951525
references.citation: Proc Natl Acad Sci U S A. 2011 May 3;108(18):7363-7
references.pmid: 21482800
references.citation: Plant Physiol. 2008 May;147(1):381-90
references.pmid: 18359842
references.citation: Trends Plant Sci. 2004 Jan;9(1):13-7
references.pmid: 14729214
references.citation: J Mol Biol. 1994 Oct 21;243(2):173-8
references.pmid: 7932747
references.citation: Cryobiology. 2005 Oct;51(2):230-4
references.pmid: 16098506
references.citation: Trends Biochem Sci. 2002 Oct;27(10):527-33
references.pmid: 12368089
references.citation: Plant Cell. 2011 Jun;23(6):2391-404
references.pmid: 21665998
references.citation: Protein Sci. 2011 Jan;20(1):42-50
references.pmid: 21031484
references.citation: Biochem Biophys Res Commun. 2005 May 27;331(1):325-32
references.pmid: 15929202
references.citation: Mol Biotechnol. 2013 Jun;54(2):643-50
references.pmid: 23054631
references.citation: Anal Biochem. 2009 Sep 1;392(1):70-6
references.pmid: 19464251
references.citation: Biochem Biophys Res Commun. 1992 Mar 31;183(3):1103-8
references.pmid: 1567390