期刊
JOURNAL OF PROTEOME RESEARCH
卷 20, 期 1, 页码 485-497出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.0c00521
关键词
programmed cell death ligand-1; PD-L1; N-glycosylation; site-specific glycosylation; polyLacNAc; immune checkpoints; therapeutic antibody; durvalumab; binding constant; surface plasmon resonance
资金
- National Institutes of Health [U01 CA207824, P41 GM103390, S10OD023557, U01 CA230692, R01CA135069, R01CA238455]
- CCSG Grant [P30 CA51008]
Immune checkpoint inhibitors, such as PD-L1/PD-1, play crucial roles in regulating immune responses and are promising targets for cancer immunotherapy. The glycosylation of PD-L1, particularly at the N219 sequon, influences its interaction with PD-1, with complex glycans and polyLacNAc structures being predominant. Different forms of PD-L1 exhibit varying N219 occupancy and polyLacNAc contributions, potentially impacting their interactions with PD-1.
Immune checkpoint inhibitors, including PD-L1/PD-1, are key regulators of the immune response and promising targets in cancer immunotherapy. N-glycosylation of PD-L1 affects its interaction with PD-1, but little is known about the distribution of glycoforms at its four NXS/T sequons. We optimized LC-MS/MS methods using collision energy modulation for the site-specific resolution of specific glycan motifs. We demonstrate that PD-L1 on the surface of breast cancer cell line carries mostly complex glycans with a high proportion of polyLacNAc structures at the N219 sequon. Contrary to the full-length protein, the secreted form of PD-L1 expressed in breast MDA-MB-231 or HEK293 cells demonstrated minimum N219 occupancy and low contribution of the polyLacNAc structures. Molecular modeling of PD-L1/PD-1 interaction with N-glycans suggests that glycans at the N219 site of PD-L1 and N74 and N116 of PD-1 may be involved in glycan-glycan interactions, but the impact of this potential interaction on the protein function remains at this point unknown. The interaction of PD-L1 with clinical antibodies is also affected by glycosylation. In conclusion, PD-L1 expressed in the MDA-MB-231 breast cancer cell line carries polyLacNAc glycans mostly at the N219 sequon, which displays the highest variability in occupancy and is most likely to influence the interaction with PD-1.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据