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The surface of<jats:italic>A. phagocytophilum<\/jats:italic>plays a crucial role in subverting the hostile host cell environment. However, except for the P44\/Msp2 outer membrane protein family, the surface components of<jats:italic>A. phagocytophilum<\/jats:italic>are largely unknown. To identify the major surface proteins of<jats:italic>A. phagocytophilum<\/jats:italic>, a membrane-impermeable, cleavable biotin reagent, sulfosuccinimidyl-2-[biotinamido]ethyl-1,3-dithiopropionate (Sulfo-NHS-SS-Biotin), was used to label intact bacteria. The biotinylated bacterial surface proteins were isolated by streptavidin agarose affinity purification and then separated by electrophoresis, followed by capillary liquid chromatography-nanospray tandem mass spectrometry analysis. Among the major proteins captured by affinity purification were five<jats:italic>A. phagocytophilum<\/jats:italic>proteins, Omp85, hypothetical proteins APH_0404 (designated Asp62) and APH_0405 (designated Asp55), P44 family proteins, and Omp-1A. The surface exposure of Asp62 and Asp55 was verified by immunofluorescence microscopy. Recombinant Asp62 and Asp55 proteins were recognized by an HGA patient serum. Anti-Asp62 and anti-Asp55 peptide sera partially neutralized<jats:italic>A. phagocytophilum<\/jats:italic>infection of HL-60 cells in vitro. We found that the Asp62 and Asp55 genes were cotranscribed and conserved among members of the family<jats:italic>Anaplasmataceae<\/jats:italic>. With the exception of P44-18, all of the proteins were newly revealed major surface-exposed proteins whose study should facilitate understanding the interaction between<jats:italic>A. phagocytophilum<\/jats:italic>and the host. These proteins may serve as targets for development of chemotherapy, diagnostics, and vaccines.<\/jats:p>","DOI":"10.1128\/jb.00866-07","type":"journal-article","created":{"date-parts":[[2007,9,1]],"date-time":"2007-09-01T00:53:59Z","timestamp":1188608039000},"page":"7819-7828","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":57,"title":["Identification of Novel Surface Proteins of<i>Anaplasma phagocytophilum<\/i>by Affinity Purification and Proteomics"],"prefix":"10.1128","volume":"189","author":[{"given":"Yan","family":"Ge","sequence":"first","affiliation":[{"name":"Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio 43210"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuko","family":"Rikihisa","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio 43210"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"1322","DOI":"10.3201\/eid1108.050085","volume":"11","year":"2005","unstructured":"Alberti, A., M. 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