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Characterization of the ice-binding protein from Arctic yeast Leucosporidium sp. AY30

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Title: Characterization of the ice-binding protein from Arctic yeast Leucosporidium sp. AY30
Authors: Park, Kyoung Sun1,2, Do, Hackwon1,2, Lee, Jun Hyuck1,2, Park, Seung Il3, Kim, Eun jung1, Kim, Soon-Jong4, Kang, Sung-Ho1,2, Kim, Hak Jun1,2 hjkim@kopri.re.kr
Source: Cryobiology. Jun2012, Vol. 64 Issue 3, p286-296. 11p.
Subjects: Antifreeze proteins, Chromatographic analysis, Ultracentrifugation, Green fluorescent protein, Circular dichroism, Glycosylation
Abstract: Abstract: Previously, we reported the ice-binding protein (LeIBP) from the Arctic yeast Leucosporidium sp. AY30. In this study we provide physicochemical characterization of this IBP, which belongs to a class of IBPs that exhibited no significant similarity in primary structure to other known antifreeze proteins (AFPs). We compared native, glycosylated and non-glycosylated recombinant LeIBPs. Interestingly, size-exclusion chromatography and analytical ultracentrifugation revealed that LeIBP self-associates with a reversible dimer with K d values in the range 3.45–7.24×10−6 M. Circular dichroism (CD) spectra showed that LeIBP, glycosylated or non-glycosylated, is predominantly composed of β-strand secondary structural elements (54.6%), similar to other β-helical antifreeze proteins (AFPs). In thermal hysteresis (TH) activity measurements, native LeIBP was twice more active (0.87°C at 15mg/mL) than that of the recombinant IBPs (0.43–0.42°C at 10.8mg/mL). This discrepancy is probably due to uncharacterized enhancing factors carried over during ice affinity purification, because glycosylated and non-glycosylated recombinant proteins displayed similarly low activity. Ice recrystallization inhibition (RI) activities of the native and recombinant LeIBPs were comparable. Measurements of CD, TH activity, and RI showed that glycosylation does not cause structural changes and is not required for function. An ice-etching experiment using green fluorescent protein-tagged IBP revealed that LeIBP binds, just as hyperactive AFPs, to both basal and pyramidal prism planes of the ice crystal. Taken together, our results indicate that LeIBP, structurally similar to hyperactive AFPs, is moderately active and that a reversible dimer has no effect on its activity. [Copyright &y& Elsevier]
Copyright of Cryobiology is the property of Academic Press Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Characterization of the ice-binding protein from Arctic yeast Leucosporidium sp. AY30
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  Data: <searchLink fieldCode="AR" term="%22Park%2C+Kyoung+Sun%22">Park, Kyoung Sun</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Do%2C+Hackwon%22">Do, Hackwon</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Lee%2C+Jun+Hyuck%22">Lee, Jun Hyuck</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Park%2C+Seung+Il%22">Park, Seung Il</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Kim%2C+Eun+jung%22">Kim, Eun jung</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Kim%2C+Soon-Jong%22">Kim, Soon-Jong</searchLink><relatesTo>4</relatesTo><br /><searchLink fieldCode="AR" term="%22Kang%2C+Sung-Ho%22">Kang, Sung-Ho</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Kim%2C+Hak+Jun%22">Kim, Hak Jun</searchLink><relatesTo>1,2</relatesTo><i> hjkim@kopri.re.kr</i>
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  Data: <searchLink fieldCode="JN" term="%22Cryobiology%22">Cryobiology</searchLink>. Jun2012, Vol. 64 Issue 3, p286-296. 11p.
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  Data: <searchLink fieldCode="DE" term="%22Antifreeze+proteins%22">Antifreeze proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Chromatographic+analysis%22">Chromatographic analysis</searchLink><br /><searchLink fieldCode="DE" term="%22Ultracentrifugation%22">Ultracentrifugation</searchLink><br /><searchLink fieldCode="DE" term="%22Green+fluorescent+protein%22">Green fluorescent protein</searchLink><br /><searchLink fieldCode="DE" term="%22Circular+dichroism%22">Circular dichroism</searchLink><br /><searchLink fieldCode="DE" term="%22Glycosylation%22">Glycosylation</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Abstract: Previously, we reported the ice-binding protein (LeIBP) from the Arctic yeast Leucosporidium sp. AY30. In this study we provide physicochemical characterization of this IBP, which belongs to a class of IBPs that exhibited no significant similarity in primary structure to other known antifreeze proteins (AFPs). We compared native, glycosylated and non-glycosylated recombinant LeIBPs. Interestingly, size-exclusion chromatography and analytical ultracentrifugation revealed that LeIBP self-associates with a reversible dimer with K d values in the range 3.45–7.24×10−6 M. Circular dichroism (CD) spectra showed that LeIBP, glycosylated or non-glycosylated, is predominantly composed of β-strand secondary structural elements (54.6%), similar to other β-helical antifreeze proteins (AFPs). In thermal hysteresis (TH) activity measurements, native LeIBP was twice more active (0.87°C at 15mg/mL) than that of the recombinant IBPs (0.43–0.42°C at 10.8mg/mL). This discrepancy is probably due to uncharacterized enhancing factors carried over during ice affinity purification, because glycosylated and non-glycosylated recombinant proteins displayed similarly low activity. Ice recrystallization inhibition (RI) activities of the native and recombinant LeIBPs were comparable. Measurements of CD, TH activity, and RI showed that glycosylation does not cause structural changes and is not required for function. An ice-etching experiment using green fluorescent protein-tagged IBP revealed that LeIBP binds, just as hyperactive AFPs, to both basal and pyramidal prism planes of the ice crystal. Taken together, our results indicate that LeIBP, structurally similar to hyperactive AFPs, is moderately active and that a reversible dimer has no effect on its activity. [Copyright &y& Elsevier]
– Name: AbstractSuppliedCopyright
  Label:
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  Data: <i>Copyright of Cryobiology is the property of Academic Press Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1016/j.cryobiol.2012.02.014
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        Text: English
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      – SubjectFull: Ultracentrifugation
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      – SubjectFull: Circular dichroism
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      – SubjectFull: Glycosylation
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      – TitleFull: Characterization of the ice-binding protein from Arctic yeast Leucosporidium sp. AY30
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              Text: Jun2012
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