Refolding, purification, and crystallization of apical membrane antigen 1 from Plasmodium falciparum

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• 作者：Gupta ; Aditi ; Bai ; Tao ; Murphy ; Vince ; Strike ; Phillip ; Anders ; Robin F. ; Batchelor ; Adrian H.
• 关键词：Malaria vaccine refolding purification crystallization
• 刊名：Protein Expression and Purification
• 出版年：2005
• 出版时间：May, 2005
• 年：2005
• 卷：41
• 期：1
• 页码：186-198
• 全文大小：726 K

文摘

Extracellular domains of malaria antigens almost invariably contain disulphide linkages but lack N- and O-linked glycosylation. The best practical approach to generating recombinant extracellular Plasmodium proteins is not established and the problems encountered when using a bacterial expression/refolding approach are discussed in detail. Limited proteolysis experiments were used to identify a relatively non-flexible core region of the Plasmodium falciparum protein apical membrane antigen 1 (AMA1), and refolding/purification was used to generate two fragments of AMA1. Several chromatographically distinct AMA1 variants were identified that are presumably differentially refolded proteins. One of these AMA1 preparations proved to be crystallizable and generated two crystal forms that diffracted X-rays to 2 resolution.