Activation of antifreeze proteins from larvae of the beetle Dendroides canadensis |
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Authors: | Ding Wen Wu John G Duman |
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Affiliation: | (1) Department of Biological Sciences, University of Notre Dame, 46556 Notre Dame, IN, USA |
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Abstract: | Summary Purified antifreeze proteins (AFPs) from the larvae of the beetle Dendroides canadensis do not produce the high levels of antifreeze activity seen in the hemolymph of overwintering larvae, even when the purified AFPs are assayed at very high concentrations. However, addition of certain proteins or agar (at concentrations sufficiently low that the gel state does not result) to the Dendroides AFP resulted in a 2–3-fold increase in activity. A 70-kDa protein with AFP-activating capabilities was purified from Dendroides larvae. Addition of this endogenous activator protein to a 4 mg·ml-1 solution of AFP increased the activity of the AFPs to values comparable to those of the hemolymph of overwintering larvae. Data derived from a modified immunoblot technique demonstrate that the activators bind to the AFP, or vice versa. Formation of this association must allow the AFP to block ice crystal growth by binding to the surface of potential seed crystals in the normal fashion. However, because the AFP-activator complex is much larger than the AFP alone, the complex probably blocks a greater surface area of the crystal and is thus a more efficient antifreeze.Abbreviations
AFP
antifreeze protein
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BSA
bovine serum albumine
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DEAE
diethylaminoethyl
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Ig
immunoglubolin
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LPIN
lipoprotein ice nucleator
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PIN
protein ice nucleator
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SDS
sodium dodecyl sulfate
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PAGE
polyacrylamide gel electrophoresis
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TH
thermal hysteresis |
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Keywords: | Antitreeze proteins Cold tolerance Thermal hysteresis proteins Insect cold tolerance |
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