Structural and biochemical studies of HCMV gH/gL/gO and Pentamer reveal mutually exclusive cell entry complexes

被引：124

作者：

Ciferri, Claudio ^{[1
]}

Chandramouli, Sumana ^{[1
]}

Donnarumma, Danilo ^{[2
]}

Nikitin, Pavel A. ^{[3
]}

Cianfrocco, Michael A. ^{[4
,5
]}

Gerrein, Rachel ^{[1
]}

Feire, Adam L. ^{[3
]}

Barnett, Susan W. ^{[1
]}

Lilja, Anders E. ^{[1
]}

Rappuoli, Rino ^{[2
]}

Norais, Nathalie ^{[2
]}

Settembre, Ethan C. ^{[1
]}

Carfi, Andrea ^{[1
]}

机构：

[1] Novartis Vaccines, Cambridge, MA 02139 USA

[2] Novartis Vaccines, I-53100 Siena, Italy

[3] Novartis Inst Biomed Res, Emeryville, CA 94608 USA

[4] Harvard Univ, Dept Mol & Cellular Biol, Cambridge, MA 02138 USA

[5] Harvard Univ, Sch Med, Dept Cell Biol, Boston, MA 02115 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2015年 / 112卷 / 06期

关键词：

human cytomegalovirus; HCMV; Pentamer complex; gH/gL/gO; virus entry; GLYCOPROTEIN-H; CRYSTAL-STRUCTURE; NEUTRALIZING ANTIBODIES; CYTOMEGALOVIRUS DISEASE; PSEUDORABIES VIRUS; ENDOTHELIAL-CELLS; AMINO-TERMINUS; IN-VITRO; FUSION; GH;

D O I：

10.1073/pnas.1424818112

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Human cytomegalovirus (HCMV) is a major cause of morbidity and mortality in transplant patients and the leading viral cause of birth defects after congenital infection. The glycoprotein complexes gH/gL/gO and gH/gL/UL128/UL130/UL131A (Pentamer) are key targets of the human humoral response against HCMV and are required for HCMV entry into fibroblasts and endothelial/epithelial cells, respectively. We expressed and characterized soluble forms of gH/gL, gH/gL/gO, and Pentamer. Mass spectrometry and mutagenesis analysis revealed that gL-Cys144 forms disulfide bonds with gO-Cys351 in gH/gL/gO and with UL128-Cys162 in the Pentamer. Notably, Pentamer harboring the UL128-Cys162Ser/gL-Cys144Ser mutations had impaired syncytia formation and reduced interference of HCMV entry into epithelial cells. Electron microscopy analysis showed that HCMV gH/gL resembles HSV gH/gL and that gO and UL128/UL130/UL131A bind to the same site at the gH/gL N terminus. These data are consistent with gH/gL/gO and Pentamer forming mutually exclusive cell entry complexes and reveal the overall location of gH/gL-, gH/gL/gO-, and Pentamer-specific neutralizing antibody binding sites. Our results provide, to our knowledge, the first structural view of gH/gL/gO and Pentamer supporting the development of vaccines and antibody therapeutics against HCMV.

引用

页码：1767 / 1772

页数：6

共 39 条

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