2001 Fiscal Year Final Research Report Summary
Molecular interaction between joining chain and immunoglobulin by BIA
| Project/Area Number |
11307043
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
病態科学系歯学(含放射線系歯学)
|
|---|
| Research Institution | Nihon-University |
|---|
Principal Investigator |
MORO Itaru Nihon-University, School of Dentistry, Professor, 歯学部, 教授 (50059531)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TAKENOBUCHI Ohkubo Nihon-University, School of Dentistry, Instructor, 歯学部, 助手 (50246914)
TAKAHASHI Tomihisa Nihon-University, School of Dentistry, Instructor, 歯学部, 助手 (40246905)
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Keywords | Mucosal immunology / Secretory IgA / Joining chain / Molecular interaction / BIA |
|---|
| Research Abstract |
Immunoglobulin (Ig) Joining (J) chain is required for polymerization of pIgs such as dimeric IgA and pentameric IgM, and has a well-conserved feature among various vertebrate and invertebrate species. Recombinant protein of chicken J chain (rCJ), molecular mass of 25 kDa, was synthesized from E. coli transformed by chicken J chain CDNA. Interaction between rCJ and Igs was analyzed by BIA CORE 3000 system. In result, rCJ has a high binding capacity to α and μ chains but not those of human. The reactivity between anti-human J chain and rCJ was much weaker than that of anti-chicken J chain. In addition, rCJ was made from cDNA substituted by cysteine residue to serine and analyzed the binding capacity to chicken Igs, resulting that no inhibitory effect was detected between α and μ chains. Chicken genomic J chain clones encoding 14 kb were taken from DNA library. The structure of its gene has 4 exons having a high degree of similarities compared to that of human and mouse. However, in 5'-region encoding 3.8 kb, there is no homologous sequence compared to mouse and cow. Luciferase assay was performed to determine the promoter activity among deletion mutants derived from 5'-region, resulting that luciferase activities were observed in all mutans. Futhermore, enhancer region consisted of 0.5 kb was isolated from 3.8 kb to 3.3 kb upstream and several franscriptional binding sites, such as NF-E2, USF-1, MyoD, CdxA and GATA3, were identified using TRANSFUC data base analysis. These results suggest that transcriptional mechanism of the chicken J chain may be different from that of mammals.
|
