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Purification of full-length recombinant human huntingtin proteins with allelic series of polyglutamine lengths

ABSTRACT: Summary Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by the polyglutamine (polyQ) expansion in huntingtin (HTT) protein. The challenge of obtaining full-length HTT proteins with high purity limits the understanding of the HTT protein function. Here, we provide a protocol to generate and purify full-length recombinant human HTT proteins with various polyQ lengths, which is key to investigate the biochemical function of HTT proteins and the molecular mechanism underlying HD pathology. For complete details on the use and execution of this protocol, please refer to Jung et al. (2020). Graphical abstract Highlights • Expression and purification of full-length recombinant human huntingtin (HTT) proteins• Generation of HTT proteins with allelic series of polyglutamine length• Purified HTT proteins form complexes with HAP40 or HAP1. Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by the polyglutamine (polyQ) expansion in huntingtin (HTT) protein. The challenge of obtaining full-length HTT proteins with high purity limits the understanding of the HTT protein function. Here, we provide a protocol to generate and purify full-length recombinant human HTT proteins with various polyQ lengths, which is key to investigate the biochemical function of HTT proteins and the molecular mechanism underlying HD pathology.

SUBMITTER: Kim H

PROVIDER: S-EPMC8551504 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective neuropathogenesis in BACHD mice.

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2017-03-22 | PXD005641 | Pride

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2017-05-26 | PXD006302 | Pride

Proteome analysis of huntingtin knock-in allelic series - Muscle

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2018-09-03 | PXD010957 | Pride

Proteome analysis of huntingtin knock-in allelic series - Heart

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