Cryo-EM Visualization of an Exposed RGD Epitope on Adenovirus That Escapes Antibody Neutralization

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1997 Mar 17

PMID 9135136

Citations 61

Authors

P L Stewart

C Y Chiu

S Huang

T Muir

Y Zhao

B Chait

P Mathias

G R Nemerow

Affiliations

Soon will be listed here.

Abstract

Interaction of the adenovirus penton base protein with alpha v integrins promotes virus entry into host cells. The location of the integrin binding sequence Arg-Gly-Asp (RGD) on human type 2 adenovirus (Ad2) was visualized by cryo-electron microscopy (cryo-EM) and image reconstruction using a mAb (DAV-1) which recognizes a linear epitope, IRGDTFATR. The sites for DAV-1 binding corresponded to the weak density above each of the five 22 A protrusions on the adenovirus penton base protein. Modeling of a Fab fragment crystal structure into the adenovirus-Fab cryo-EM density indicated a large amplitude of motion for the Fab and the RGD epitope. An unexpected finding was that Fab fragments, but not IgG antibody molecules, inhibited adenovirus infection. Steric hindrance from the adenovirus fiber and a few bound IgG molecules, as well as epitope mobility, most likely prevent binding of IgG antibodies to all five RGD sites on the penton base protein within the intact virus. These studies indicate that the structure of the adenovirus particle facilitates interaction with cell integrins, whilst restricting binding of potentially neutralizing antibodies.

Citing Articles

Integrin-Targeting Strategies for Adenovirus Gene Therapy.

Nemerow G Viruses. 2024; 16(5).

PMID: 38793651 PMC: 11125847. DOI: 10.3390/v16050770.

Biographical Feature: Charles Y. Chiu, M.D., Ph.D.

Munson E J Clin Microbiol. 2024; 62(5):e0140523.

PMID: 38619264 PMC: 11077958. DOI: 10.1128/jcm.01405-23.

Highly synergistic combinations of nanobodies that target SARS-CoV-2 and are resistant to escape.

Mast F, Fridy P, Ketaren N, Wang J, Jacobs E, Olivier J Elife. 2021; 10.

PMID: 34874007 PMC: 8651292. DOI: 10.7554/eLife.73027.

Mass spectrometric detection of KRAS protein mutations using molecular imprinting.

Norman R, Singh R, Muskett F, Parrott E, Rufini A, Langridge J Nanoscale. 2021; 13(48):20401-20411.

PMID: 34854867 PMC: 8675027. DOI: 10.1039/d1nr03180e.

Hitting the Target but Missing the Point: Recent Progress towards Adenovirus-Based Precision Virotherapies.

Cunliffe T, Bates E, Parker A Cancers (Basel). 2020; 12(11).

PMID: 33187160 PMC: 7696810. DOI: 10.3390/cancers12113327.

References

van Heel M . Angular reconstitution: a posteriori assignment of projection directions for 3D reconstruction. Ultramicroscopy. 1987; 21(2):111-23. DOI: 10.1016/0304-3991(87)90078-7. View

Smith T, Chase E, Schmidt T, Olson N, Baker T . Neutralizing antibody to human rhinovirus 14 penetrates the receptor-binding canyon. Nature. 1996; 383(6598):350-4. PMC: 4167671. DOI: 10.1038/383350a0. View

Fitzgerald D, Padmanabhan R, Pastan I, Willingham M . Adenovirus-induced release of epidermal growth factor and pseudomonas toxin into the cytosol of KB cells during receptor-mediated endocytosis. Cell. 1983; 32(2):607-17. DOI: 10.1016/0092-8674(83)90480-4. View

Adrian M, Dubochet J, Lepault J, McDowall A . Cryo-electron microscopy of viruses. Nature. 1984; 308(5954):32-6. DOI: 10.1038/308032a0. View

Seth P, Fitzgerald D, Ginsberg H, Willingham M, Pastan I . Evidence that the penton base of adenovirus is involved in potentiation of toxicity of Pseudomonas exotoxin conjugated to epidermal growth factor. Mol Cell Biol. 1984; 4(8):1528-33. PMC: 368944. DOI: 10.1128/mcb.4.8.1528-1533.1984. View

Svensson U . Role of vesicles during adenovirus 2 internalization into HeLa cells. J Virol. 1985; 55(2):442-9. PMC: 254952. DOI: 10.1128/JVI.55.2.442-449.1985. View

Seth P, Willingham M, Pastan I . Binding of adenovirus and its external proteins to Triton X-114. Dependence on pH. J Biol Chem. 1985; 260(27):14431-4. View

Wadell G, Allard A, Johansson M, Svensson L, Uhnoo I . Enteric adenoviruses. Ciba Found Symp. 1987; 128:63-91. DOI: 10.1002/9780470513460.ch5. View

Neumann R, Chroboczek J, Jacrot B . Determination of the nucleotide sequence for the penton-base gene of human adenovirus type 5. Gene. 1988; 69(1):153-7. DOI: 10.1016/0378-1119(88)90389-7. View

10.

Acharya R, Fry E, Stuart D, Fox G, Rowlands D, Brown F . The three-dimensional structure of foot-and-mouth disease virus at 2.9 A resolution. Nature. 1989; 337(6209):709-16. DOI: 10.1038/337709a0. View

11.

Parry N, Fox G, Rowlands D, Brown F, Fry E, Acharya R . Structural and serological evidence for a novel mechanism of antigenic variation in foot-and-mouth disease virus. Nature. 1990; 347(6293):569-72. DOI: 10.1038/347569a0. View

12.

Adler M, Lazarus R, Dennis M, Wagner G . Solution structure of kistrin, a potent platelet aggregation inhibitor and GP IIb-IIIa antagonist. Science. 1991; 253(5018):445-8. DOI: 10.1126/science.1862345. View

13.

Karlsson R, Michaelsson A, Mattsson L . Kinetic analysis of monoclonal antibody-antigen interactions with a new biosensor based analytical system. J Immunol Methods. 1991; 145(1-2):229-40. DOI: 10.1016/0022-1759(91)90331-9. View

14.

Main A, Harvey T, Baron M, Boyd J, Campbell I . The three-dimensional structure of the tenth type III module of fibronectin: an insight into RGD-mediated interactions. Cell. 1992; 71(4):671-8. DOI: 10.1016/0092-8674(92)90600-h. View

15.

Smith T, Olson N, Cheng R, Liu H, Chase E, Lee W . Structure of human rhinovirus complexed with Fab fragments from a neutralizing antibody. J Virol. 1993; 67(3):1148-58. PMC: 237479. DOI: 10.1128/JVI.67.3.1148-1158.1993. View

16.

Wickham T, Mathias P, Cheresh D, Nemerow G . Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell. 1993; 73(2):309-19. DOI: 10.1016/0092-8674(93)90231-e. View

17.

Stewart P, Fuller S, Burnett R . Difference imaging of adenovirus: bridging the resolution gap between X-ray crystallography and electron microscopy. EMBO J. 1993; 12(7):2589-99. PMC: 413505. DOI: 10.1002/j.1460-2075.1993.tb05919.x. View

18.

Bai M, Harfe B, Freimuth P . Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells. J Virol. 1993; 67(9):5198-205. PMC: 237917. DOI: 10.1128/JVI.67.9.5198-5205.1993. View

19.

Greber U, Willetts M, Webster P, Helenius A . Stepwise dismantling of adenovirus 2 during entry into cells. Cell. 1993; 75(3):477-86. DOI: 10.1016/0092-8674(93)90382-z. View

20.

Liu H, Smith T, Lee W, Mosser A, Rueckert R, Olson N . Structure determination of an Fab fragment that neutralizes human rhinovirus 14 and analysis of the Fab-virus complex. J Mol Biol. 1994; 240(2):127-37. DOI: 10.1006/jmbi.1994.1427. View