Protocol for detecting histidine polyphosphate modification of human proteins via MBP-tagged expression in E. coli.
STAR Protoc
; 5(2): 102947, 2024 Jun 21.
Article
em En
| MEDLINE
| ID: mdl-38470910
ABSTRACT
Polyphosphate exhibits a unique post-translational modification-like function, known as histidine polyphosphate modification (HPM), marked by a robust non-covalent interaction with histidine repeat proteins. Here, we present a protocol for detecting HPM of human proteins via maltose-binding protein-tagged expression in E. coli. We describe steps for detecting HPM by observing electrophoretic mobility shifts on NuPAGE gels followed by western blot. We then detail procedures for analyzing the influence of ionic strength and pH on HPM. For complete details on the use and execution of this protocol, please refer to Neville et al.1.
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Base de dados:
MEDLINE
Assunto principal:
Polifosfatos
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Processamento de Proteína Pós-Traducional
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Escherichia coli
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Proteínas Ligantes de Maltose
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Histidina
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article