Dissecting neurofilament tail sequence- phosphorylation-structure relationships with multicomponent reconstituted protein brushes

• 成果类型: Article
• 署名作者: Ding, Erika A.; Yokokura, Takashi J.; Wang, Rui; Kumar, Sanjay
• 署名单位: University of California System; University of California Berkeley; United States Department of Energy (DOE); Lawrence Berkeley National Laboratory; University of California System; University of California Berkeley; University of California System; University of California San Francisco
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-9261
• DOI: 10.1073/pnas.2410109121
• 发表日期: 2024-12-03
• 关键词: conformational properties; triplet proteins; nf-h; subunit; stoichiometry; crossbridges; architecture; equilibrium; simulations; morphology

摘要

Neurofilaments (NFs) are multisubunit, bottlebrush- shaped intermediate filaments abundant in the axonal cytoskeleton. Each NF subunit contains a long intrinsically disordered tail domain, which protrudes from the NF core to form a brush surrounding each NF. Precisely how the tails' variable charge patterns and repetitive phosphorylation sites mediate their conformation within the brush remains an open question in axonal biology. We address this problem by grafting recombinant NF tail protein conprotein brushes of defined stoichiometry that can be phosphorylated in vitro. Atomic force microscopy measurements reveal that brush height depends on composition monotonically but not always linearly for binary NFL:NFM or NFL:NFH systems, tail repel phosphorylated residues to generate the multilayer morphology, while the establishes a platform for dissecting contributions of disordered protein sequence to