Dataset Information

Site-Specific C-Terminal Fluorescent Labeling of Tau Protein.

ABSTRACT: Formation of Tau protein aggregates in neurons is a pathological hallmark of several neurodegenerative diseases, including Alzheimer's disease. Fluorescently labeled Tau protein is therefore useful to study the aggregation of these pathological proteins and to identify potential therapeutic targets. Conventionally, cysteine residues are used for labeling Tau proteins; however, the full-length Tau isoform contains two cysteine residues in the microtubule-binding region, which are implicated in Tau aggregation by forming intermolecular disulfide bonds. To prevent the fluorescent label from disturbing the microtubule binding region, we developed a strategy to fluorescently label Tau at its C-terminus while leaving cysteine residues unperturbed. We took advantage of a Sortase A-mediated transpeptidation approach to bind a short peptide (GGGH₆-Alexa₆₄₇) with a His-tag and a covalently attached Alexa 647 fluorophore to the C-terminus of Tau. This reaction relies on the presence of a Sortase recognition motif (LPXTG), which we attached to the C-terminus of recombinantly expressed Tau. We demonstrate that C-terminal modification of Tau protein results in no significant differences between the native and C-terminally labeled Tau monomer with regard to aggregation kinetics, secondary structure, and fibril morphology.

SUBMITTER: Bryan L

PROVIDER: S-EPMC9773802 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Publications

Site-Specific C-Terminal Fluorescent Labeling of Tau Protein.

Bryan Louise L Awasthi Saurabh S Li Yuanjie Y Nirmalraj Peter Niraj PN Balog Sandor S Yang Jerry J Mayer Michael M

ACS omega 20221212 50

Formation of Tau protein aggregates in neurons is a pathological hallmark of several neurodegenerative diseases, including Alzheimer's disease. Fluorescently labeled Tau protein is therefore useful to study the aggregation of these pathological proteins and to identify potential therapeutic targets. Conventionally, cysteine residues are used for labeling Tau proteins; however, the full-length Tau isoform contains two cysteine residues in the microtubule-binding region, which are implicated in Ta ...[more]

PMID: 36570287

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Dataset Information

Site-Specific C-Terminal Fluorescent Labeling of Tau Protein.

Publications

Site-Specific C-Terminal Fluorescent Labeling of Tau Protein.

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