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亲和纯化-质谱法分离和鉴定与人类血浆蛋白相互作用的登革病毒组。

Isolation and Identification of Dengue Virus Interactome with Human Plasma Proteins by Affinity Purification-Mass Spectrometry.

机构信息

Division of System Biology, Center for Genetic Engineering and Biotechnology, Havana, Cuba.

出版信息

Methods Mol Biol. 2022;2409:133-153. doi: 10.1007/978-1-0716-1879-0_10.

Abstract

Viral proteins evolve to benefit the interaction with host proteins during the infection and replication processes. A comprehensive understanding of virus interactome with host proteins may thus lead to the identification of molecular targets for infection inhibition. We present a procedure for isolating and identifying the dengue virus interactome with human plasma proteins. It comprises the fractionation of human plasma by anion exchange chromatography, followed by affinity purification and mass spectrometry identification of the captured proteins. This procedure was applied to the characterization of the interactions of the four serotypes of dengue virus with human plasma proteins, mediated by the domain III of the envelope protein of the virus. The resulting interactome comprises 62 proteins, six of which were validated as new direct interactions of the virus with its human host.

摘要

病毒蛋白在感染和复制过程中进化以利于与宿主蛋白相互作用。因此,全面了解病毒与宿主蛋白的相互作用组可能会鉴定出感染抑制的分子靶标。我们提出了一种分离和鉴定登革热病毒与人血浆蛋白相互作用组的程序。它包括通过阴离子交换色谱法对人血浆进行分级分离,然后通过亲和纯化和捕获蛋白的质谱鉴定。该程序应用于表征登革热病毒四个血清型与病毒包膜蛋白 III 域介导的人血浆蛋白之间的相互作用。所得的相互作用组包括 62 种蛋白质,其中 6 种被验证为病毒与其人类宿主的新的直接相互作用。

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