Proteomics

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Titin BioID - protein and peptide pull-down

ABSTRACT: Proximity proteomics has greatly advanced the analysis of native protein complexes and subcellular structures in culture, but has not been amenable to study development and disease in vivo. Here, we generated a knock-in mouse with the biotin ligase (BioID) inserted at titin’s Z-disc region to identify protein networks that connect the sarcomere to signal transduction and metabolism. Our census of the sarcomeric proteome from neonatal to adult heart and quadriceps reveals how perinatal signalling, protein homeostasis and the shift to adult energy metabolism shape the properties of striated muscle cells. Mapping biotinylation sites to sarcomere structures refined our understanding of myofilament dynamics and supports the hypothesis that myosin filaments penetrate Z-discs to dampen contraction. Extending this proof of concept study to BioID fusion proteins generated with Crispr/CAS9 in animal models recapitulating human pathology will facilitate the future analysis of molecular machines and signalling hubs in physiological, pharmacological, and disease context.

INSTRUMENT(S): Orbitrap Fusion, Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Heart, Skeletal Muscle

SUBMITTER: Marieluise Kirchner  

LAB HEAD: Michael Gotthardt

PROVIDER: PXD017341 | Pride | 2020-05-18

REPOSITORIES: Pride

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Dataset's files

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Action DRS
201701_uniprot_10090_Mus_musculus_Titin.fasta Fasta
ANAKIN_20170523_MK_CF_H_NE_19100.raw Raw
ANAKIN_20170523_MK_CF_H_NE_19101.raw Raw
ANAKIN_20170523_MK_CF_H_NE_19102.raw Raw
ANAKIN_20170523_MK_CF_H_NE_19165.raw Raw
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Publications

Proximity proteomics has greatly advanced the analysis of native protein complexes and subcellular structures in culture, but has not been amenable to study development and disease in vivo. Here, we have generated a knock-in mouse with the biotin ligase (BioID) inserted at titin's Z-disc region to identify protein networks that connect the sarcomere to signal transduction and metabolism. Our census of the sarcomeric proteome from neonatal to adult heart and quadriceps reveals how perinatal signa  ...[more]

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