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タイトルSingle-Molecule 3D Images of "Hole-Hole" IgG1 Homodimers by Individual-Particle Electron Tomography.
ジャーナル・号・ページSci Rep, Vol. 9, Issue 1, Page 8864, Year 2019
掲載日2019年6月20日
著者Dongsheng Lei / Jianfang Liu / Hongbin Liu / Thomas E Cleveland / John P Marino / Ming Lei / Gang Ren /
PubMed 要旨The engineering of immunoglobulin-G molecules (IgGs) is of wide interest for improving therapeutics, for example by modulating the activity or multiplexing the specificity of IgGs to recognize more ...The engineering of immunoglobulin-G molecules (IgGs) is of wide interest for improving therapeutics, for example by modulating the activity or multiplexing the specificity of IgGs to recognize more than one antigen. Optimization of engineered IgG requires knowledge of three-dimensional (3D) structure of synthetic IgG. However, due to flexible nature of the molecules, their structural characterization is challenging. Here, we use our reported individual-particle electron tomography (IPET) method with optimized negative-staining (OpNS) for direct 3D reconstruction of individual IgG hole-hole homodimer molecules. The hole-hole homodimer is an undesired variant generated during the production of a bispecific antibody using the knob-into-hole heterodimer technology. A total of 64 IPET 3D density maps at ~15 Å resolutions were reconstructed from 64 individual molecules, revealing 64 unique conformations. In addition to the known Y-shaped conformation, we also observed an unusual X-shaped conformation. The 3D structure of the X-shaped conformation contributes to our understanding of the structural details of the interaction between two heavy chains in the Fc domain. The IPET approach, as an orthogonal technique to characterize the 3D structure of therapeutic antibodies, provides insight into the 3D structural variety and dynamics of heterogeneous IgG molecules.
リンクSci Rep / PubMed:31221961 / PubMed Central
手法EM (トモグラフィー)
解像度11.93 - 14.26 Å
構造データ

EMDB-7353:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 001)
手法: EM (トモグラフィー) / 解像度: 13.21 Å

EMDB-7354:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 002)
手法: EM (トモグラフィー) / 解像度: 13.63 Å

EMDB-7355:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 003)
手法: EM (トモグラフィー) / 解像度: 12.52 Å

EMDB-7356:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 004)
手法: EM (トモグラフィー) / 解像度: 12.81 Å

EMDB-7357:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 005)
手法: EM (トモグラフィー) / 解像度: 12.67 Å

EMDB-7358:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 006)
手法: EM (トモグラフィー) / 解像度: 14.1 Å

EMDB-7359:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 007)
手法: EM (トモグラフィー) / 解像度: 12.78 Å

EMDB-7360:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 008)
手法: EM (トモグラフィー) / 解像度: 12.88 Å

EMDB-7361:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 009)
手法: EM (トモグラフィー) / 解像度: 14.06 Å

EMDB-7362:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 010)
手法: EM (トモグラフィー) / 解像度: 13.5 Å

EMDB-7363:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 011)
手法: EM (トモグラフィー) / 解像度: 13.49 Å

EMDB-7364:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 012)
手法: EM (トモグラフィー) / 解像度: 13.34 Å

EMDB-7365:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 013)
手法: EM (トモグラフィー) / 解像度: 13.28 Å

EMDB-7366:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 014)
手法: EM (トモグラフィー) / 解像度: 13.23 Å

EMDB-7367:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 015)
手法: EM (トモグラフィー) / 解像度: 12.31 Å

EMDB-7368:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 016)
手法: EM (トモグラフィー) / 解像度: 12.88 Å

EMDB-7369:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 017)
手法: EM (トモグラフィー) / 解像度: 13.71 Å

EMDB-7370:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 018)
手法: EM (トモグラフィー) / 解像度: 13.24 Å

EMDB-7371:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 019)
手法: EM (トモグラフィー) / 解像度: 12.48 Å

EMDB-7372:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 020)
手法: EM (トモグラフィー) / 解像度: 13.93 Å

EMDB-7373:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 021)
手法: EM (トモグラフィー) / 解像度: 13.63 Å

EMDB-7374:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 022)
手法: EM (トモグラフィー) / 解像度: 13.36 Å

EMDB-7375:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 023)
手法: EM (トモグラフィー) / 解像度: 12.81 Å

EMDB-7376:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 024)
手法: EM (トモグラフィー) / 解像度: 12.63 Å

EMDB-7377:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 025)
手法: EM (トモグラフィー) / 解像度: 12.7 Å

EMDB-7378:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 026)
手法: EM (トモグラフィー) / 解像度: 14.08 Å

EMDB-7379:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 027)
手法: EM (トモグラフィー) / 解像度: 12.75 Å

EMDB-7380:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 028)
手法: EM (トモグラフィー) / 解像度: 11.93 Å

EMDB-7381:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 029)
手法: EM (トモグラフィー) / 解像度: 13.54 Å

EMDB-7382:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 030)
手法: EM (トモグラフィー) / 解像度: 13 Å

EMDB-7383:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 031)
手法: EM (トモグラフィー) / 解像度: 12.54 Å

EMDB-7384:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 032)
手法: EM (トモグラフィー) / 解像度: 13.5 Å

EMDB-7385:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 033)
手法: EM (トモグラフィー) / 解像度: 12.28 Å

EMDB-7386:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 034)
手法: EM (トモグラフィー) / 解像度: 13.7 Å

EMDB-7387:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 035)
手法: EM (トモグラフィー) / 解像度: 12.86 Å

EMDB-7388:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 036)
手法: EM (トモグラフィー) / 解像度: 11.95 Å

EMDB-7389:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 037)
手法: EM (トモグラフィー) / 解像度: 12.22 Å

EMDB-7390:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 038)
手法: EM (トモグラフィー) / 解像度: 12.26 Å

EMDB-7391:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 039)
手法: EM (トモグラフィー) / 解像度: 13.62 Å

EMDB-7392:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 040)
手法: EM (トモグラフィー) / 解像度: 12.15 Å

EMDB-7393:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 041)
手法: EM (トモグラフィー) / 解像度: 12.29 Å

EMDB-7394:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 042)
手法: EM (トモグラフィー) / 解像度: 12.36 Å

EMDB-7395:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 043)
手法: EM (トモグラフィー) / 解像度: 13.63 Å

EMDB-7396:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 044)
手法: EM (トモグラフィー) / 解像度: 13.08 Å

EMDB-7397:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 045)
手法: EM (トモグラフィー) / 解像度: 12.59 Å

EMDB-7398:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 046)
手法: EM (トモグラフィー) / 解像度: 12.58 Å

EMDB-7399:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 047)
手法: EM (トモグラフィー) / 解像度: 12.2 Å

EMDB-7400:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 048)
手法: EM (トモグラフィー) / 解像度: 13.81 Å

EMDB-7401:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 049)
手法: EM (トモグラフィー) / 解像度: 13.17 Å

EMDB-7402:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 050)
手法: EM (トモグラフィー) / 解像度: 11.93 Å

EMDB-7404:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 051)
手法: EM (トモグラフィー) / 解像度: 13.6 Å

EMDB-7405:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 052)
手法: EM (トモグラフィー) / 解像度: 12.09 Å

EMDB-7406:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 053)
手法: EM (トモグラフィー) / 解像度: 12.12 Å

EMDB-7407:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 054)
手法: EM (トモグラフィー) / 解像度: 13.88 Å

EMDB-7408:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 055)
手法: EM (トモグラフィー) / 解像度: 13.57 Å

EMDB-7409:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 056)
手法: EM (トモグラフィー) / 解像度: 13.38 Å

EMDB-7410:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 057)
手法: EM (トモグラフィー) / 解像度: 12.48 Å

EMDB-7411:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 058)
手法: EM (トモグラフィー) / 解像度: 12.31 Å

EMDB-7412:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 059)
手法: EM (トモグラフィー) / 解像度: 12.64 Å

EMDB-7413:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 060)
手法: EM (トモグラフィー) / 解像度: 12 Å

EMDB-7414:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 061)
手法: EM (トモグラフィー) / 解像度: 12.18 Å

EMDB-7415:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 062)
手法: EM (トモグラフィー) / 解像度: 14.26 Å

EMDB-7416:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 063)
手法: EM (トモグラフィー) / 解像度: 12.58 Å

EMDB-7417:
Single Molecule 3D Image of a Hole-Hole Homodimer Species Generated in the Production of a Bispecific Recombinant IgG1 (No. 064)
手法: EM (トモグラフィー) / 解像度: 12.59 Å

EMDB-7418:
Single-Molecule 3D Image of Half IgG1 (No. 001)
手法: EM (トモグラフィー) / 解像度: 12.76 Å

EMDB-7419:
Single-Molecule 3D Image of Half IgG1 (No. 002)
手法: EM (トモグラフィー) / 解像度: 13.3 Å

EMDB-7420:
Single-Molecule 3D Image of Half IgG1 (No. 003)
手法: EM (トモグラフィー) / 解像度: 13.55 Å

EMDB-7421:
Single-Molecule 3D Image of Half IgG1 (No. 004)
手法: EM (トモグラフィー) / 解像度: 12.98 Å

EMDB-7422:
Single-Molecule 3D Image of Half IgG1 (No. 005)
手法: EM (トモグラフィー) / 解像度: 12.26 Å

EMDB-7423:
Single-Molecule 3D Image of Half IgG1 (No. 006)
手法: EM (トモグラフィー) / 解像度: 12.77 Å

EMDB-7424:
Single-Molecule 3D Image of Half IgG1 (No. 007)
手法: EM (トモグラフィー) / 解像度: 13.23 Å

EMDB-7425:
Single-Molecule 3D Image of Half IgG1 (No. 008)
手法: EM (トモグラフィー) / 解像度: 12.79 Å

EMDB-7426:
Single-Molecule 3D Image of Half IgG1 (No. 009)
手法: EM (トモグラフィー) / 解像度: 12.63 Å

EMDB-7427:
Single-Molecule 3D Image of Half IgG1 (No. 010)
手法: EM (トモグラフィー) / 解像度: 14.07 Å

EMDB-7428:
Single-Molecule 3D Image of Half IgG1 (No. 011)
手法: EM (トモグラフィー) / 解像度: 13.37 Å

EMDB-7429:
Single-Molecule 3D Image of Half IgG1 (No. 012)
手法: EM (トモグラフィー) / 解像度: 13.29 Å

EMDB-7430:
Single-Molecule 3D Image of Half IgG1 (No. 013)
手法: EM (トモグラフィー) / 解像度: 12.7 Å

EMDB-7431:
Single-Molecule 3D Image of Half IgG1 (No. 014)
手法: EM (トモグラフィー) / 解像度: 13.73 Å

EMDB-7432:
Single-Molecule 3D Image of Half IgG1 (No. 015)
手法: EM (トモグラフィー) / 解像度: 13.43 Å

EMDB-7433:
Single-Molecule 3D Image of Half IgG1 (No. 016)
手法: EM (トモグラフィー) / 解像度: 13.88 Å

由来
  • unidentified (未定義)
キーワードAntibodies, Bispecific (抗体) / Electron Microscope Tomography / Imaging, Three-Dimensional / Immunoglobulin G (免疫グロブリンG) / Molecular Imaging / Negative Staining / Protein Conformation / Protein Multimerization

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