Prolyl 4-hydroxylation of alpha-fibrinogen: a novel protein modification revealed by plasma proteomics.
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.
Ono
Masaya
M
Chemotherapy Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan. masono@ncc.go.jp
Matsubara
Junichi
J
Honda
Kazufumi
K
Sakuma
Tomohiro
T
Hashiguchi
Tomoyo
T
Nose
Hiroshi
H
Nakamori
Shoji
S
Okusaka
Takuji
T
Kosuge
Tomoo
T
Sata
Naohiro
N
Nagai
Hideo
H
Ioka
Tatsuya
T
Tanaka
Sachiko
S
Tsuchida
Akihiko
A
Aoki
Tatsuya
T
Shimahara
Masashi
M
Yasunami
Yohichi
Y
Itoi
Takao
T
Moriyasu
Fuminori
F
Negishi
Ayako
A
Kuwabara
Hideya
H
Shoji
Ayako
A
Hirohashi
Setsuo
S
Yamada
Tesshi
T
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
D001798:Blood Proteins; D020543:Proteome; D005340:Fibrinogen; D011393:Procollagen-Proline Dioxygenase
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