{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,30]],"date-time":"2026-08-30T02:48:24Z","timestamp":1788058104941,"version":"build-2784847793"},"posted":{"date-parts":[[2024,5,21]]},"group-title":"Biochemistry","reference-count":43,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,5,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Glycosylphosphatidylinositol (GPI) anchor protein modification in\n                  <jats:italic>Plasmodium<\/jats:italic>\n                  species is well known and represents the principal form of glycosylation in these organisms. The structure and biosynthesis of GPI anchors of\n                  <jats:italic>Plasmodium<\/jats:italic>\n                  spp. has been primarily studied in the asexual blood stage of\n                  <jats:italic>P. falciparum<\/jats:italic>\n                  and is known to contain the typical conserved GPI structure of EtN-P-Man3GlcN-PI. Here, we have investigated the circumsporozoite protein (CSP) for the presence of a GPI-anchor. CSP is the major surface protein of\n                  <jats:italic>Plasmodium<\/jats:italic>\n                  sporozoites, the infective stage of the malaria parasite. While it is widely assumed that CSP is a GPI-anchored cell surface protein, compelling biochemical evidence for this supposition is absent. Here, we employed metabolic labeling and mass-spectrometry based approaches to confirm the presence of a GPI anchor in CSP.\n                <\/jats:p>\n                <jats:p>\n                  Biosynthetic radiolabeling of CSP with [\n                  <jats:sup>3<\/jats:sup>\n                  H]-palmitic acid and [\n                  <jats:sup>3<\/jats:sup>\n                  H]-ethanolamine, with the former being base-labile and therefore ester-linked, provided strong evidence for the presence of a GPI anchor on CSP, but these data alone were not definitive. To provide further evidence, immunoprecipitated CSP was analyzed for presence of\n                  <jats:italic>myo<\/jats:italic>\n                  -inositol (a characteristic component of GPI anchor) using strong acid hydrolysis and GC-MS for a highly sensitive and quantitative detection. The single ion monitoring (SIM) method for GC-MS analysis confirmed the presence of the\n                  <jats:italic>myo<\/jats:italic>\n                  -inositol component in CSP. Taken together, these data provide confidence that the long-assumed presence of a GPI anchor on this important parasite protein is correct.\n                <\/jats:p>","DOI":"10.1101\/2024.05.21.595204","type":"posted-content","created":{"date-parts":[[2024,5,22]],"date-time":"2024-05-22T00:20:21Z","timestamp":1716337221000},"source":"Crossref","is-referenced-by-count":1,"title":["The major surface protein of malaria sporozoites is GPI-anchored to the plasma membrane"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2127-3115","authenticated-orcid":false,"given":"Rupa","family":"Nagar","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefano S.","family":"Garcia Castillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Pinzon-Ortiz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sharon","family":"Patray","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alida","family":"Coppi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sachie","family":"Kanatani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert L.","family":"Moritz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristian E.","family":"Swearingen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael A. J.","family":"Ferguson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Photini","family":"Sinnis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2024052407001067000_2024.05.21.595204v1.1","doi-asserted-by":"publisher","DOI":"10.1101\/GLYCOBIOLOGY.4E.12"},{"key":"2024052407001067000_2024.05.21.595204v1.2","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PPAT.1005259"},{"key":"2024052407001067000_2024.05.21.595204v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2023.12.006"},{"key":"2024052407001067000_2024.05.21.595204v1.4","first-page":"2597","article-title":"Glycosylphosphatidylinositols synthesized by asexual erythrocytic stages of the malarial parasite, Plasmodium falciparum","volume":"269","year":"1994","journal-title":"Candidates for plasmodial glycosylphosphatidylinositol membrane anchor precursors and pathogenicity factors. J. 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