Antigenic characterization of an intrinsically unstructured protein, plasmodium falciparum merozoite surface protein 2

Christopher G Adda, Christopher Andrew MacRaild, Linda Reiling, Kaye Wycherley, Michelle J Boyle, Vivian Kienzle, Paul Masendycz, Michael Foley, James G Beeson, Raymond Stanley Norton, Robin Anders

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Merozoite surface protein 2 (MSP2) is an abundant glycosylphosphatidylinositol (GPI)-anchored protein of Plasmodium falciparum, which is a potential component of a malaria vaccine. As all forms of MSP2 can be categorized into two allelic families, a vaccine containing two representative forms of MSP2 may overcome the problem of diversity in this highly polymorphic protein. Monomeric recombinant MSP2 is an intrinsically unstructured protein, but its conformational properties on the merozoite surface are unknown. This question is addressed here by analyzing the 3D7 and FC27 forms of recombinant and parasite MSP2 using a panel of monoclonal antibodies raised against recombinant MSP2. The epitopes of all antibodies, mapped using both a peptide array and by nuclear magnetic resonance (NMR) spectroscopy on full-length recombinant MSP2, were shown to be linear. The antibodies revealed antigenic differences, which indicate that the conserved N- and C-terminal regions, but not the central variable region, are less accessible in the parasite antigen. This appears to be an intrinsic property of parasite MSP2 and is not dependent on interactions with other merozoite surface proteins as the loss of some conserved-region epitopes seen using the immunofluorescence assay (IFA) on parasite smears was also seen on Western blot analyses of parasite lysates. Further studies of the structural basis of these antigenic differences are required in order to optimize recombinant MSP2 constructs being evaluated as potential vaccine components.
Original languageEnglish
Pages (from-to)4177 - 4185
Number of pages9
JournalInfection and Immunity
Volume80
Issue number12
DOIs
Publication statusPublished - 2012

Cite this

Adda, Christopher G ; MacRaild, Christopher Andrew ; Reiling, Linda ; Wycherley, Kaye ; Boyle, Michelle J ; Kienzle, Vivian ; Masendycz, Paul ; Foley, Michael ; Beeson, James G ; Norton, Raymond Stanley ; Anders, Robin. / Antigenic characterization of an intrinsically unstructured protein, plasmodium falciparum merozoite surface protein 2. In: Infection and Immunity. 2012 ; Vol. 80, No. 12. pp. 4177 - 4185.
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abstract = "Merozoite surface protein 2 (MSP2) is an abundant glycosylphosphatidylinositol (GPI)-anchored protein of Plasmodium falciparum, which is a potential component of a malaria vaccine. As all forms of MSP2 can be categorized into two allelic families, a vaccine containing two representative forms of MSP2 may overcome the problem of diversity in this highly polymorphic protein. Monomeric recombinant MSP2 is an intrinsically unstructured protein, but its conformational properties on the merozoite surface are unknown. This question is addressed here by analyzing the 3D7 and FC27 forms of recombinant and parasite MSP2 using a panel of monoclonal antibodies raised against recombinant MSP2. The epitopes of all antibodies, mapped using both a peptide array and by nuclear magnetic resonance (NMR) spectroscopy on full-length recombinant MSP2, were shown to be linear. The antibodies revealed antigenic differences, which indicate that the conserved N- and C-terminal regions, but not the central variable region, are less accessible in the parasite antigen. This appears to be an intrinsic property of parasite MSP2 and is not dependent on interactions with other merozoite surface proteins as the loss of some conserved-region epitopes seen using the immunofluorescence assay (IFA) on parasite smears was also seen on Western blot analyses of parasite lysates. Further studies of the structural basis of these antigenic differences are required in order to optimize recombinant MSP2 constructs being evaluated as potential vaccine components.",
author = "Adda, {Christopher G} and MacRaild, {Christopher Andrew} and Linda Reiling and Kaye Wycherley and Boyle, {Michelle J} and Vivian Kienzle and Paul Masendycz and Michael Foley and Beeson, {James G} and Norton, {Raymond Stanley} and Robin Anders",
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Antigenic characterization of an intrinsically unstructured protein, plasmodium falciparum merozoite surface protein 2. / Adda, Christopher G; MacRaild, Christopher Andrew; Reiling, Linda; Wycherley, Kaye; Boyle, Michelle J; Kienzle, Vivian; Masendycz, Paul; Foley, Michael; Beeson, James G; Norton, Raymond Stanley; Anders, Robin.

In: Infection and Immunity, Vol. 80, No. 12, 2012, p. 4177 - 4185.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Antigenic characterization of an intrinsically unstructured protein, plasmodium falciparum merozoite surface protein 2

AU - Adda, Christopher G

AU - MacRaild, Christopher Andrew

AU - Reiling, Linda

AU - Wycherley, Kaye

AU - Boyle, Michelle J

AU - Kienzle, Vivian

AU - Masendycz, Paul

AU - Foley, Michael

AU - Beeson, James G

AU - Norton, Raymond Stanley

AU - Anders, Robin

PY - 2012

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N2 - Merozoite surface protein 2 (MSP2) is an abundant glycosylphosphatidylinositol (GPI)-anchored protein of Plasmodium falciparum, which is a potential component of a malaria vaccine. As all forms of MSP2 can be categorized into two allelic families, a vaccine containing two representative forms of MSP2 may overcome the problem of diversity in this highly polymorphic protein. Monomeric recombinant MSP2 is an intrinsically unstructured protein, but its conformational properties on the merozoite surface are unknown. This question is addressed here by analyzing the 3D7 and FC27 forms of recombinant and parasite MSP2 using a panel of monoclonal antibodies raised against recombinant MSP2. The epitopes of all antibodies, mapped using both a peptide array and by nuclear magnetic resonance (NMR) spectroscopy on full-length recombinant MSP2, were shown to be linear. The antibodies revealed antigenic differences, which indicate that the conserved N- and C-terminal regions, but not the central variable region, are less accessible in the parasite antigen. This appears to be an intrinsic property of parasite MSP2 and is not dependent on interactions with other merozoite surface proteins as the loss of some conserved-region epitopes seen using the immunofluorescence assay (IFA) on parasite smears was also seen on Western blot analyses of parasite lysates. Further studies of the structural basis of these antigenic differences are required in order to optimize recombinant MSP2 constructs being evaluated as potential vaccine components.

AB - Merozoite surface protein 2 (MSP2) is an abundant glycosylphosphatidylinositol (GPI)-anchored protein of Plasmodium falciparum, which is a potential component of a malaria vaccine. As all forms of MSP2 can be categorized into two allelic families, a vaccine containing two representative forms of MSP2 may overcome the problem of diversity in this highly polymorphic protein. Monomeric recombinant MSP2 is an intrinsically unstructured protein, but its conformational properties on the merozoite surface are unknown. This question is addressed here by analyzing the 3D7 and FC27 forms of recombinant and parasite MSP2 using a panel of monoclonal antibodies raised against recombinant MSP2. The epitopes of all antibodies, mapped using both a peptide array and by nuclear magnetic resonance (NMR) spectroscopy on full-length recombinant MSP2, were shown to be linear. The antibodies revealed antigenic differences, which indicate that the conserved N- and C-terminal regions, but not the central variable region, are less accessible in the parasite antigen. This appears to be an intrinsic property of parasite MSP2 and is not dependent on interactions with other merozoite surface proteins as the loss of some conserved-region epitopes seen using the immunofluorescence assay (IFA) on parasite smears was also seen on Western blot analyses of parasite lysates. Further studies of the structural basis of these antigenic differences are required in order to optimize recombinant MSP2 constructs being evaluated as potential vaccine components.

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DO - 10.1128/IAI.00665-12

M3 - Article

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