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Protocol for detecting histidine polyphosphate modification of human proteins via MBP-tagged expression in E. coli.

Kirsten Lehotsky | Nolan Neville | Zongchao Jia
STAR protocols | 2024

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.

Pubmed ID: 38470910

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This is a list of tools and resources that we have found mentioned in this publication.


Millipore (tool)

RRID:SCR_008983

An Antibody supplier

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MBP (8G1) Mouse mAb (antibody)

RRID:AB_2140060

This monoclonal targets MBL2

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MBP (8G1) Mouse mAb (antibody)

RRID:AB_2140060

This monoclonal targets MBL2

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