دورية أكاديمية
CryoEM structure of a therapeutic antibody (favezelimab) bound to human LAG3 determined using a bivalent Fab as fiducial marker.
| العنوان: | CryoEM structure of a therapeutic antibody (favezelimab) bound to human LAG3 determined using a bivalent Fab as fiducial marker. |
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| المؤلفون: | Mishra AK; W.M. Keck Laboratory for Structural Biology, University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA., Shahid S; W.M. Keck Laboratory for Structural Biology, University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA., Karade SS; W.M. Keck Laboratory for Structural Biology, University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA., Agnihotri P; W.M. Keck Laboratory for Structural Biology, University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA., Kolesnikov A; W.M. Keck Laboratory for Structural Biology, University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA., Hasan SS; Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD 21201, USA; University of Maryland Marlene and Stewart Greenebaum Cancer Center, University of Maryland Medical Center, Baltimore, MD 21201, USA; Center for Biomolecular Therapeutics, University of Maryland School of Medicine, Rockville, MD 20850, USA., Mariuzza RA; W.M. Keck Laboratory for Structural Biology, University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA. Electronic address: rmariuzz@umd.edu. |
| المصدر: | Structure (London, England : 1993) [Structure] 2023 Oct 05; Vol. 31 (10), pp. 1149-1157.e3. Date of Electronic Publication: 2023 Aug 23. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Research Support, N.I.H., Extramural |
| اللغة: | English |
| بيانات الدورية: | Publisher: Cell Press Country of Publication: United States NLM ID: 101087697 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1878-4186 (Electronic) Linking ISSN: 09692126 NLM ISO Abbreviation: Structure Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2000- : Cambridge, Mass. : Cell Press Original Publication: London : Current Biology, c1993- |
| مواضيع طبية MeSH: | Fiducial Markers* , Antibodies, Monoclonal*/metabolism, Humans ; Cryoelectron Microscopy/methods ; T-Lymphocytes/metabolism ; Binding Sites |
| مستخلص: | Lymphocyte activation gene 3 protein (LAG3) is an inhibitory receptor that is upregulated on exhausted T cells in tumors. LAG3 is a major target for cancer immunotherapy with many anti-LAG3 antibodies in clinical trials. However, there is no structural information on the epitopes recognized by these antibodies. We determined the single-particle cryoEM structure of a therapeutic antibody (favezelimab) bound to LAG3 to 3.5 Å resolution, revealing that favezelimab targets the LAG3-binding site for MHC class II, its canonical ligand. The small size of the complex between the conventional (monovalent) Fab of favezelimab and LAG3 (∼100 kDa) presented a challenge for cryoEM. Accordingly, we engineered a bivalent version of Fab favezelimab that doubled the size of the Fab-LAG3 complex and conferred a highly identifiable shape to the complex that facilitated particle selection and orientation for image processing. This study establishes bivalent Fabs as new fiducial markers for cryoEM analysis of small proteins. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 Elsevier Ltd. All rights reserved.) |
| References: | Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67. (PMID: 21460454) Curr Opin Struct Biol. 2009 Feb;19(1):39-49. (PMID: 19162470) J Mol Biol. 2021 Jan 22;433(2):166714. (PMID: 33220264) Cancer Res. 2014 Jul 1;74(13):3418-28. (PMID: 24769443) Nat Struct Mol Biol. 2019 Jun;26(6):510-517. (PMID: 31160781) Bioinformatics. 2003 Jul 1;19(10):1290-1. (PMID: 12835274) Acta Crystallogr D Biol Crystallogr. 2004 Mar;60(Pt 3):432-8. (PMID: 14993666) J Immunol. 2022 Oct 15;209(8):1586-1594. (PMID: 36104110) J Immunol Methods. 2018 May;456:7-14. (PMID: 29427592) Acta Crystallogr D Biol Crystallogr. 1996 May 1;52(Pt 3):534-42. (PMID: 15299676) Proc Natl Acad Sci U S A. 2021 Oct 12;118(41):. (PMID: 34620716) Cell. 2019 Jan 10;176(1-2):334-347.e12. (PMID: 30580966) N Engl J Med. 2022 Jan 6;386(1):24-34. (PMID: 34986285) Proc Natl Acad Sci U S A. 1997 May 27;94(11):5744-9. (PMID: 9159144) Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3. (PMID: 15299374) Cancer Cell. 2015 Apr 13;27(4):450-61. (PMID: 25858804) J Exp Med. 1990 May 1;171(5):1393-405. (PMID: 1692078) N Engl J Med. 2010 Aug 19;363(8):711-23. (PMID: 20525992) Eur J Immunol. 2021 Feb;51(2):331-341. (PMID: 32920841) Nat Methods. 2017 Mar;14(3):290-296. (PMID: 28165473) Nature. 2020 Mar;579(7799):443-447. (PMID: 32103179) Science. 2016 Sep 30;353(6307):. (PMID: 27708076) Nat Immunol. 2019 Nov;20(11):1425-1434. (PMID: 31611702) Structure. 2012 Apr 4;20(4):582-92. (PMID: 22483106) Proc Natl Acad Sci U S A. 2021 Nov 23;118(47):. (PMID: 34782475) Clin Cancer Res. 2022 Dec 1;28(23):5030-5039. (PMID: 35579997) Nature. 2019 Dec;576(7786):315-320. (PMID: 31776516) Nat Struct Mol Biol. 2018 Mar;25(3):289-296. (PMID: 29434346) Protein Sci. 2002 Jun;11(6):1285-99. (PMID: 12021428) J Mol Biol. 2018 Feb 2;430(3):337-347. (PMID: 29273204) Immunooncol Technol. 2022 Mar 17;14:100079. (PMID: 35755891) Nat Immunol. 2018 Dec;19(12):1415-1426. (PMID: 30349037) J Mol Biol. 2007 Sep 21;372(3):774-97. (PMID: 17681537) Cancer Immunol Res. 2022 Oct 4;10(10):1175-1189. (PMID: 35981087) Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501. (PMID: 20383002) N Engl J Med. 2019 Oct 17;381(16):1535-1546. (PMID: 31562797) Nat Immunol. 2022 Jul;23(7):1031-1041. (PMID: 35761082) Acta Crystallogr D Biol Crystallogr. 2012 Apr;68(Pt 4):352-67. (PMID: 22505256) Acta Crystallogr D Biol Crystallogr. 2006 Aug;62(Pt 8):859-66. (PMID: 16855301) Elife. 2018 May 24;7:. (PMID: 29792401) Nature. 1988 Sep 8;335(6186):188-90. (PMID: 3412476) Immunity. 2016 May 17;44(5):989-1004. (PMID: 27192565) Cancer Res. 2012 Feb 15;72(4):917-27. (PMID: 22186141) |
| معلومات مُعتمدة: | P30 GM138396 United States GM NIGMS NIH HHS; R01 AI144422 United States AI NIAID NIH HHS; P30 CA134274 United States CA NCI NIH HHS; HHSN261200800001E United States CA NCI NIH HHS |
| فهرسة مساهمة: | Keywords: LAG3; T cell; X-ray crystallography; antibody; cryoEM; fiducial marker; inhibitory receptor |
| المشرفين على المادة: | 0 (Antibodies, Monoclonal) |
| تواريخ الأحداث: | Date Created: 20230824 Date Completed: 20231009 Latest Revision: 20240627 |
| رمز التحديث: | 20240627 |
| مُعرف محوري في PubMed: | PMC11197462 |
| DOI: | 10.1016/j.str.2023.07.013 |
| PMID: | 37619561 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1878-4186 |
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| DOI: | 10.1016/j.str.2023.07.013 |