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1998 Fiscal Year Final Research Report Summary

Host defense mechanism and modulation of lipid metabolism by pulmonary surfactant proteins

Research Project

Project/Area Number 09670159
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Pathological medical chemistry
Research InstitutionSapporo Medical University

Principal Investigator

KUROKI Yoshio  Sapporo Medical University, School of Medicine, Professor, 医学部, 教授 (70161784)

Co-Investigator(Kenkyū-buntansha) SANO Hitomi  Sapporo Medical University, School of Medicine, Research Associate, 医学部, 助手 (80295344)
SOHMA Hitoshi  Sapporo Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (70226702)
Project Period (FY) 1997 – 1998
Keywordspulmonary surfactant / SP-A / SP-D / mannose-binding protein / collectin / lipopolysaccharides / CD14 / TNFalpha
Research Abstract

1. Pulmonary surfactant proteins A and D (SP-A and SP-D) and mannose-binding protein A(MBP-A) are collectins in the C-type lectin superfamily. These collectins exhibit unique lipid binding properties. The structure-function relationship was investigated by using SP-ASP-D chimeras, It was found that the SP-A region of Glu^<195>- Phe^<228> is required for lipid and type II cell interactions and that the SP-D region of Cys^<261>-Phe^<355> is required for optimal lipid interactions.
Monoclonal antibodies (mAbs PElO and PC6) that recognize human SP-A inhibit the interactions of SP-A with lipids and alveolar type II cells. We mapped the epitopes for antihuman SP-A mAbs by a phage display peptide library. Phage selected by mAbs displayed the consensus peptide sequences that are nearly identical to ^<184>TPVNYTNWYRG^<194> of human SP-A.Chimeric proteins were generated in which the rat SP-A region Thr^<174>-Gly^<194> was replaced with the MBP-A region Thr^<164>-Asp^<184> (rat ama4, respectively) … More . Rat ama4 bound to an affinity matrix on mannose-sepharose but lost all of the SP-A functions except carbohydrate binding and Ca^<2+> independent GalCer binding. Strikingly, the rat ama4 chimera acquired the PI binding property that MBP-A exhibits. This study demonstrates that the amino acid residues 174-194 of SP-A and the corresponding region of MBP-A are critical for SP-A-type II cell interaction and Ca^<2+>-dependent lipid binding of collectins.
2. Pulmonary surfactant protein A (SP-A) plays an important part in antibody independent host defense mechanisms of the lung. SP-A bound to rough forms but not to smooth forms of LPS.When U937 cells and rat alveolar macrophages were preincubated with SP-A ; smooth LPS failed to induce TNFalpha secretion while rough LPS-induced TNFalpha secretion was modestly increased. Western blot analysis revealed that CD14 was one of the SP-A-binding proteins isolated from solubilized U937 cells. In addition, SP-A directly bound to recombinant soluble CD14 (rsCDl4). When rsCDl4 was preincubated with SP-A, the binding of rsCDl4 to smooth LPS was significantly reduced but the association of rsCDl4 with rough LPS was augmented. These results demonstrate the different actions of SP-A upon distinct serotypes of LPS, and indicate that the direct interaction of SP-A with CD14 constitutes a likely mechanism by which SP-A modulates LPS-elicited cellular responses. Less

  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Honma T: "The mannose-binding protein A region of Glu185-Ala221 can functionally replace the surfactant protein A region of Glu195-Phe228 without loss of interaction with lipidsand alveolar type II cells." Biochemistry. 36. 7176-7184 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hattori A: "IgG is associated with surfactant protein A aggregate isolated from alveolar proteinosis patients" AmJ Respir Crit Care Med. 156. 1785-1788 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kuroki Y: FEBS Lett. 414. 387-392 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagae H: "Enzyme-linked immunosorbent assay using F(ab')2 fragment for the detection of human pulmonary surfactant protein D in sera." Clin Chem Acta.266. 157-171 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 黒木由夫: "肺サーファクタントの生化学-アポ蛋白質の構造と機能の解析" 呼吸と循環. 45. 533-541 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sano H: "Analysis of chimeric proteins identified the regions in the carbohydrate recgnition domain of rat lung collectins that are essential for interactions with phospholipids,glycolipids and alyeolar type IIcells." J Biol Chem. 273. 4783-4789 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsunezawa W: "Site-directed mutagenesis of surfactant protein A reveals dissociation of lipid aggregation and lipid uptake by alveolar type II cells." Biochim Biophys Acta. 1387. 433-446 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Pattanajitvilai S: "Mutational analysis of Arg197 of rat surfactant protein A.His197 creates specific lipid uptake defects." J Biol Chem. 273. 5702-5707 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shijubo N: "BAL surfactant protein A and Clara cells 10 kDa protein levels in healthy subjects." Lung. 176. 257-265 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kuroki Y: "Surfactant proteins A and D:disease markers." Biochim Biophys Acta. 1408. 334-345 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mason RJ: "A 50 kDa variant form of human surfactant protein D." Eur Respir J. 12. 1147-1155 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tanaka T: "Molecular modeling of single polypeptide chain of calcium-binding protein p26olf from dimeric S100B." Protein Eng. (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 黒木由夫: "肺胞表面をおおうサーファクタント-特異蛋白質の構造と機能の解析" 蛋白質核酸酵素. 43. 834-846 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sohma H: "Quantitative Reduction of Type I Adenylyl Cyclase in Human Alcoholics." Biochim Biophys Acta. (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sano H: "Pulmonary surfactant protein A modulates the cellular response to smooth and rough lipopolysaccharides by interaction with CD14." J Immunol. (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sohma H: "Characterization of the Ca2+-dependent binding of annexin IV to surfactant protein A." Biochem J. (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chiba H: "Induction of mannose binding protein-type phosphatidylinositol recognition into pulmonary surfactant protein A." Biochemistry. (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 黒木由夫: "肺サーファクタント蛋白質SP-AおよびSP-D." 日本臨床. (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 秋野豊明: "呼吸器疾患の分子生物学:脂質代謝" 川上義和,谷口直之,木田厚瑞, 447 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 黒木由夫: "呼吸器疾患の分子生物学:サーファクタントの産生と代謝" 川上義和,谷口直之,木田厚瑞, 447 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Honma T: "The mannose-binding protein Aregion of Glu^<185>-Ala^<221> can functionally replace the surfactant protein A region of Glu^<195>-Phe^<228> without loss of interaction with lipidsand alveolar type II cells." Biochemistry. 36. 7176-7184 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hattori A: "IgG is associated with surfactant protein A aggregate isolated from alveolar proteinosis patients." Am J Respir Crit Care Med. 155. 1785-1788 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kuroki Y: "A novel type of binding specificity to phospholipids for rat mannose binding proteins isolated from serum and liver." FEBS Lett. 414. 387-392 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagae H: "Enzyme-linked immunosorbent assay using F (ab') 2 fragment for the detectin of human pulmonary surfactant protein D." Clin Chem Acta. 266. 157-171 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sano H: "Analysis of chimeric proteins identified the regions in the carbohydrate recognition domain of rat lung collectins that are essential for interactions with phospholipids, glycolipids and alveolar type II cells." J Biol Chem. 273. 4783-4789 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsunezawa W: "Site-directed mutagenesis of surfactant portein A reveals dissociation of lipid aggregatin and lipid uptake by alveolar type II cells." Biochim Biophys Acta. 1387. 433-466 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Pattanajitvilai S: "Mutational analysis of Arg197 of rat surfactant protein A.His197 creates specific lipid uptake defect." J Biol Chem. 273. 5702-5707 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shijubo N: "BAL surfactant protein A and Clara cells 10 kDa protein levels in healthy subjects." Lung. 176. 257-265 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kuroki Y: "Surfactant proteins A and D : diseases markers." Biochim Biophys Acta. 1408. 334-345 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mason RJ: "A 50 kDa variant form of human surfactant protein D." Eur Respir J. 12. 1147-1155 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tanaka T: "Molecular modeling of single polypeptide chain of calcium-binding protein p26olf from dimeric S100B." Protein Eng. (in press.). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sohma H: "Quantitative reduction of type I adenylyl cyclase in human alcoholics." Biochim Biophys Acta. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sano H: "Pulmonary surfactant protein A modulates the cellular response to smooth and rough lipopolysaccharides by intertaction with CD14" J Immunol. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sohma H: "Characterization of the Ca^<2+>-dependent binding of annexin IV to surfactant protein A." Biochem J. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chiba H: "Introduction of mannose binding protein-type phosphatidylinositol recognition into pulmonary surfactant protein A." Biochemistry. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-12-08  

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