Structural Analysis of Serum Ceruloplasmin Peptides From The Patients With Wilson disease and Familial Hypoccerulosminemia

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17591125
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Kurume University
• Principal Investigator: WATANABE Yoriko Kurume University, School of Medicine, Assistant Professor (40258489)
• Co-Investigator(Kenkyū-buntansha): YOSHINO Makoto Kurume University, School of Medicine, Professor (40080569) ; HARADA Eimei Kurume University, School of medicine, Assistant (90309790)
• Project Period (FY): 2005 – 2007
• Keywords: Cerulonlasmin / proteome analysis / hypoceruloplasminemia / Wilson disease / MALDI-TOF-MS

Research Abstract

The ceruloplasmin (Cp)-IgG complexes were extracted from plasma of the patient with hypoceruloplasminemia (genotype: nonsense mutation/unknown) and healthy controls, and were subjected to the two dimensional isoelectric focusing (IEF) analysis. Expected smaller peptide molecules due to the nonsense mutation in patient with hypoceruloplasminemia were not identified. This IEF analysis was also performed with plasma from the patient with Wilson disease who had hypoceruloproteinemia and no ceruloplasmin gene mutations. This study also did not show any patient's specific bands. After excision of the gel pieces containing the bands from the patient with hypoceruloplasminemia, the patient with Wilson disease and controls. They are subjected to analysis by an AXIMA-CFR Plus (Shimadzu / Kratos Analytical) matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) equipped with a delayed extraction and operated at a 20 kV accelerating voltage in a reflector and the positive ion mode. This analysis did not reveal any specific peaks suggestive of disease specific molecular weight. This might be due to a low sensitivity in identifying peptide peaks by this method. Detectability of the standard ceruloplasmin peptide peaks was found to be approximately 19%. This might explain the reason why no distinct peaks reflecting the gene mutations were identified by this method. We found no specific changes in IEF analysis and MALDI-TOF-MS analysis in the patient with Wilson disease. This might be due to lacking post translational modification in ceruloplasmin synthesis or due to that the changes outside of the range of detectable peptide peaks. Based on the fact that the standard human ceruloplasmin peptide peaks were detectable by only 19% by MALDI-TOF-MS analysis, we believe that MALDI-TOF-MS has limited value for protein structure analysis.

Research Products

• [Journal Article] Paternal transmission and slow elimination of mutant alleles associated with late-onset ornithine transcarbamylase deficiency in male patients (2008)
  • Author(s): Numata S, Harada E, Maeno Y, Ueki I, Watanabe Y, Fujii C, Yanagawa T, Takenaka S, Inoue T, Inoue S, Goushi T, Yasutake T, Mizuta T, Yoshino M
  • Journal Title: J Hum Genet 53 Pages: 10-17
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Paternal transmission and slow elimination of mutant alleles associated with late-onset ornithine transcarbamylase deficiency in male patients. (2008)
  • Author(s): Numata S, Harada E, Maeno Y, Ueki I, Watanabe Y, Fujii C, Yanagawa T, Takenaka S, Inoue T, Inoue S, Goushi T, Yasutake T, Mizuta T, Yoshino M.
  • Journal Title: J Hum Genet 53(doi ; 10.1007/s10038-007-0212-8) Pages: 10-17
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Roles of specific cytokines in bone remodeling and hematopoiesis in Gaucher disease (2007)
  • Author(s): Yoshino M, Watanabe Y, Tokunaga Y, Harada E, Fujii C, Numata S, Harada M, Tajima A, Ida H
  • Journal Title: Pediatr Int 49 Pages: 959-965
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Novel mutations in five Japanese patients with 3-methylcrotonyl-CoA carboxylase deficiency (2007)
  • Author(s): Uematsu M, Sakamoto O, Sugawara N, Kumagai N, Morimoto T, Yamaguchi S, Hasegawa Y, Kobayashi H, Ihara K, Yoshino M, Watanabe Y, Inokuchi T, Yokoyama T, Kiwaki K, Nakamura K, Endo F, Tsuchiya S, Ohura T
  • Journal Title: J Hum Genet 52 Pages: 1040-1043
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Screening for Alagille syndrome mutations in the JAG1 and NOTCH2 genes using denaturing high-performance liquid chromatography (2007)
  • Author(s): Samejima H, Torii C, Kosaki R, Kurosawa K, Yoshihashi H, Muroya K, Okamoto N, Watanabe Y, Kosho T, Kubota M, Matsuda O, Goto M, Izumi K, Takahashi T, Kosaki K
  • Journal Title: Genet Test 11 Pages: 216-227
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Roles of some cytokines in bone remodeling and hematopoiesis in Gaucher disease. (2007)
  • Author(s): Yoshino M, Watanabe Y, Thkunaga Y, Harada E, Fujii C, Numata S, Harada M, Tajima A, Ida H.
  • Journal Title: Pediatr Int 49(doi : 10.1111/j.1442-200x.2007.02502.x) Pages: 959-956
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Novel mutations in five Japanese patients with 3-methylcrotonyl-C0A carboxylase deficiency. (2007)
  • Author(s): Uematsu M, Sakamoto O, Sugawara N, Kumagai N, Morimoto T,Yamaguchi S, Hasegawa Y, Kobayashi H, Ihara K, Yoshino M,Watanabe Y, Inokuchi T, Yokoyama T. Kiwaki K, Nakamura K, Endo F, Tsuchiya S, Ohura T.
  • Journal Title: J Hum Genet. 52(doi : 10.1007/s10038-007-0211-9) Pages: 1040-1043
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Screening for Alagille syndrome mutations in the JAG1 and NOTCH2 genes using denaturing high-performance liquid chromatography. (2007)
  • Author(s): Samejima H,'Ibrii C, Kosaki R, Kurosawa K, Yoshihashi H,Muroya K, Okamoto N, Watanabe Y, Kosho T, Kubota M, Matsuda O, Goto M, Izumi K, Takahashi'F, Kosaki K.
  • Journal Title: Genet Test 11(doi : 10.1089/gte.2006.0519) Pages: 216-227
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Late-onset ornithine transcarbamylase deficiency in male patients:Prognostic factors and characteristics of plasma amino acid profile (2006)
  • Author(s): Harada E, Nishiyori A, Tokunaga Y, Watanabe Y, Kuriya N, Kumashiro R, Kuno T, Kuromaru R, Hirose S, Ichikawa K, Yoshino M
  • Journal Title: Pediatr Int 48 Pages: 105-111
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Comprehensive genetic analysis of relevant four genes in 49 patients with Marfan syndrome or Marfan-related phenotypes (2006)
  • Author(s): Sakai H, Visser R, Ikegawa S, Ito E, Numabe H, Watanabe Y, Mikami H, Kondoh T, Kitoh H, Sugiyama R, Okamoto N, Ogata T, Fodde R, Mizuno S, Takamura K, Egashira M, Sasaki N, Watanabe S, Nishimaki S, Takada F, Nagai T, Okada Y, Aoka Y, Yasuda K, Iwasa M, Kogaki S, Harada N, Mizuguchi T, Matsumoto N
  • Journal Title: Am J Med Genet A 140・16 Pages: 1719-1725
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Late-onset ornithine transcarbamylase deficiency inmale patients:Prognostic factors and characteristics of plasma amino acid profile. (2006)
  • Author(s): Harada E, Nishiyori A, Tokunaga Y, Watanabe Y, Kuriya N,Kumasbiro R, Kuno T, Kuromaru R, Hirose 5, Ichikawa K,Yoshino M.
  • Journal Title: Pediatr Int 48(doi : 10.1111/j.1442-200x.02181.x) Pages: 105-111
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Comprehensive genetic analysis of relevant fourgenes in 49 patients with Marfan syndrome or Marfan-related phenotypes. (2006)
  • Author(s): Sakai H, Visser R, Ikegawa S, Ito E, Numabe H, Watanabe Y, Mikami H, Kondoh T, Kitoh H, Sugiyama R, Okamoto N, Ogata T, Fodde R, Mizuno S, Takamura K, Egashira M, Sasaki N,Watanabe S, Nishimaki S, Takada F, Nagai T, Okada Y, Aoka Y, Yasuda K, Iwasa M, Kogaki S, Harada N, Mizuguchi T, Matsumoto N.
  • Journal Title: Am J Med GenetA. 140(16)(doi : 10.1002/ajmg.a.31353) Pages: 1719-25
  • Description: 「研究成果報告書概要(欧文)」より