title: Prolyl 4-hydroxylation of alpha-fibrinogen: a novel protein modification revealed by plasma proteomics.
abstract: Plasma proteome analysis requires sufficient power to compare numerous samples and detect changes in protein modification, because the protein content of human samples varies significantly among individuals, and many plasma proteins undergo changes in the bloodstream. A label-free proteomics platform developed in our laboratory, termed "Two-Dimensional Image Converted Analysis of Liquid chromatography and mass spectrometry (2DICAL)," is capable of these tasks. Here, we describe successful detection of novel prolyl hydroxylation of alpha-fibrinogen using 2DICAL, based on comparison of plasma samples of 38 pancreatic cancer patients and 39 healthy subjects. Using a newly generated monoclonal antibody 11A5, we confirmed the increase in prolyl-hydroxylated alpha-fibrinogen plasma levels and identified prolyl 4-hydroxylase A1 as a key enzyme for the modification. Competitive enzyme-linked immunosorbent assay of 685 blood samples revealed dynamic changes in prolyl-hydroxylated alpha-fibrinogen plasma level depending on clinical status. Prolyl-hydroxylated alpha-fibrinogen is presumably controlled by multiple biological mechanisms, which remain to be clarified in future studies.
authors.affiliation: Chemotherapy Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan. masono@ncc.go.jp
authors.forename: Masaya
authors.initials: M
authors.lastname: Ono
authors.forename: Junichi
authors.initials: J
authors.lastname: Matsubara
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Honda
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Sakuma
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Hashiguchi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nose
authors.forename: Shoji
authors.initials: S
authors.lastname: Nakamori
authors.forename: Takuji
authors.initials: T
authors.lastname: Okusaka
authors.forename: Tomoo
authors.initials: T
authors.lastname: Kosuge
authors.forename: Naohiro
authors.initials: N
authors.lastname: Sata
authors.forename: Hideo
authors.initials: H
authors.lastname: Nagai
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Ioka
authors.forename: Sachiko
authors.initials: S
authors.lastname: Tanaka
authors.forename: Akihiko
authors.initials: A
authors.lastname: Tsuchida
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Aoki
authors.forename: Masashi
authors.initials: M
authors.lastname: Shimahara
authors.forename: Yohichi
authors.initials: Y
authors.lastname: Yasunami
authors.forename: Takao
authors.initials: T
authors.lastname: Itoi
authors.forename: Fuminori
authors.initials: F
authors.lastname: Moriyasu
authors.forename: Ayako
authors.initials: A
authors.lastname: Negishi
authors.forename: Hideya
authors.initials: H
authors.lastname: Kuwabara
authors.forename: Ayako
authors.initials: A
authors.lastname: Shoji
authors.forename: Setsuo
authors.initials: S
authors.lastname: Hirohashi
authors.forename: Tesshi
authors.initials: T
authors.lastname: Yamada
mesh_terms: D000230:Adenocarcinoma / Q000097:blood; D001798:Blood Proteins / Q000737:chemistry*; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005340:Fibrinogen / Q000737:chemistry*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D013058:Mass Spectrometry / Q000379:methods; D010190:Pancreatic Neoplasms / Q000097:blood; D011393:Procollagen-Proline Dioxygenase / Q000737:chemistry*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D034622:RNA Interference
chemical_list: D001798:Blood Proteins; D020543:Proteome; D005340:Fibrinogen; D011393:Procollagen-Proline Dioxygenase
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