ASP53, a thermostable protein from Acacia erioloba seeds that protects target proteins against thermal denaturation
Linda Mtwisha A , Jill M. Farrant A , Wolf Brandt A , Caswell Hlongwane B and George G. Lindsey A CA Department of Molecular and Cell Biology, University of Cape Town, Private Bag 7725, Rondebosch, South Africa.
B CSIR Bio/Chemtek, P.O. Box 395, Pretoria, South Africa.
C Corresponding author. Email: George.Lindsey@uct.ac.za
Functional Plant Biology 34(2) 139-149 https://doi.org/10.1071/FP06135
Submitted: 25 May 2006 Accepted: 18 January 2007 Published: 12 February 2007
Abstract
ASP53, a 53 kDa heat soluble protein, was identified as the most abundant protein in the mature seeds of Acacia erioloba E.Mey. Immunocytochemistry showed that ASP53 was present in the vacuoles and cell walls of the axes and cotyledons of mature seeds and disappeared coincident with loss of desiccation tolerance. The sequence of the ASP53 transcript was determined and found to be homologous to the double cupin domain-containing vicilin class of seed storage proteins. Mature seeds survived heating to 60°C and this may be facilitated by the presence of ASP53. Circular dichroism spectroscopy demonstrated that the protein displayed defined secondary structure, which was maintained even at high temperature. ASP53 was found to inhibit all three stages of protein thermal denaturation. ASP53 decreased the rate of loss of alcohol dehydrogenase activity at 55°C, decreased the rate of temperature-dependent loss of secondary structure of haemoglobin and completely inhibited the temperature-dependent aggregation of egg white protein.
Additional keywords: circular dichroism spectroscopy, immunocytochemistry, protection of protein conformation, seed storage protein.
Acknowledgements
GL and JMF acknowledge the support of the National Research Foundation and the UCT Research Fund.
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