Dataset Information

PTMC_A pyridinium-based strategy for bioorthogonal conjugation-assisted purification method for profiling cell surface proteome

ABSTRACT: Cell surface proteins (CSPs) are valuable targets for therapeutic agents, but achieving highly selective CSP enrichment in cellular physiology remains a technical challenge. To address this challenge, we propose a newly developed sulfo-pyridinium-ester (SPE) cross-linking probe, followed by two-step imaging and enrichment. The SPE probe shows higher efficiency in labelling proteins at the level of cell lysates than similar NHS esters and demonstrates specificity for Lys in competitive experiments. More importantly, this probe can selectively label cell membranes in cell imaging, while only negligible labelling of the intracellular compartment was detected. Moreover, we successfully performed this strategy to mainly label the cell surface proteins in MCF-7 live cells. Finally, we used our probe to label CSPs under insulin treatment, revealing several cell surface targets for key functional biomarkers and the pathogenesis of insulin-stimulated MCF-7 breast cancer cells. The above results show that the SPE method provides a promising tool for selectively labelling cell surface proteins and monitoring transient cell surface events.

ORGANISM(S): Homo Sapiens

SUBMITTER: Rui Wang

PROVIDER: PXD044541 | iProX | Fri Aug 11 00:00:00 BST 2023

REPOSITORIES: iProX

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Publications

Pyridinium-Based Strategy for a Bioorthogonal Conjugation-Assisted Purification Method for Profiling Cell Surface Proteome.

Dai Chuan C Guo Xiaochun X Pan Zhuoheng Z Wan Chuan C Yang Dongyan D Li Yongli Y Lian Chenshan C An Yuhao Y Zhang Tuanjie T Yang Fadeng F Zhu Lizhi L Yin Feng F Wang Rui R Li Zigang Z

Analytical chemistry 20231107 46

Cell surface proteins (CSPs) are valuable targets for therapeutic agents, but achieving highly selective CSP enrichment in cellular physiology remains a technical challenge. To address this challenge, we propose a newly developed sulfo-pyridinium ester (SPE) cross-linking probe, followed by two-step imaging and enrichment. The SPE probe showed higher efficiency in labeling proteins than similar NHS esters at the level of cell lysates and demonstrated specificity for Lys in competitive experiment ...[more]

PMID: 37934015

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Project description:Background: Mortality from preeclampsia (PE) and PE-associated morbidities are 3-to 5-fold higher in persons of African ancestry than in those of Asian and European ancestries. The placenta is central to the etiology of PE. However, how and to what extent the placenta contributes to worse PE outcomes in persons of African ancestry is yet to be fully elucidated. Objective: We aimed to identify molecular pathways that are unique or enriched in placentas of parturient persons of African ancestry with PE with severe features (sPE) compared to those of Asian and European ancestry with sPE. Study design: Bulk RNA sequencing was performed on 50 placentas from parturient persons with sPE of African (n=9), Asian (n=18) and European (n=23) ancestries and 73 normotensive controls of African (n=9), Asian (n=15) and European (n=49) ancestries. Results: Metabolism, hormone regulation and hypoxia/angiogenesis genes, previously described to be upregulated in PE, including: LEP, PAPPA2, INHA, FSTL3, FLT1, PHYHIP and ENG, were upregulated in sPE across ancestries, with high expression of FSTL3 being additionally associated with intrauterine growth restriction (p<.0001). Notably, LEP, FLT1 and PHYHIP were more highly upregulated in sPE placentas from parturient persons of African versus Asian ancestry. Genes associated with allograft rejection and adaptive immune response were selectively upregulated in placentas from African ancestry parturitions and not in those of Asian and European ancestries. Among the allograft rejection/adaptive immune response genes, IL3RA was of particular interest because the patient with the highest placental IL3RA level, a woman of African ancestry with IL3RA levels 4.5-fold above the average for African ancestry parturitions with sPE, developed postpartum cardiomyopathy, and was the only patient out of 123, that developed this condition. Interestingly, the sPE patients of Asian and European ancestries with the highest IL3RA levels for their ancestries developed unexplained tachycardia peripartum, necessitating echocardiography in the European ancestry patient. The association between elevated placental IL3RA levels and unexplained tachycardia or peripartum cardiomyopathy was found to be significant in the 50 sPE patients (p=.0005). Conclusions: African ancestry parturitions are associated with higher placental upregulation of metabolism (LEP) and hypoxia/angiogenesis (FLT1) genes, and selective upregulation of allograft rejection/adaptive immune response genes, including IL3RA. High placental expression of IL3RA may predict worse maternal cardiovascular outcomes, including peripartum cardiomyopathy. Studies evaluating placental IL3RA levels in peripartum cardiomyopathy cohorts are therefore warranted, as are broader studies evaluating placental factors in maternal cardiovascular outcomes postpartum.

Dataset Information

PTMC_A pyridinium-based strategy for bioorthogonal conjugation-assisted purification method for profiling cell surface proteome

Publications

Pyridinium-Based Strategy for a Bioorthogonal Conjugation-Assisted Purification Method for Profiling Cell Surface Proteome.

Similar Datasets

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