Measuring expression heterogeneity of single-cell cytoskeletal protein complexes

Julea Vlassakis, Louise L. Hansen, Ryo Higuchi-Sanabria, Yun Zhou, C. Kimberly Tsui, Andrew Dillin, Haiyan Huang and Amy E. Herr ()
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Julea Vlassakis: University of California Berkeley
Louise L. Hansen: University of California Berkeley
Ryo Higuchi-Sanabria: University of California Berkeley
Yun Zhou: University of California Berkeley
C. Kimberly Tsui: University of California Berkeley
Andrew Dillin: University of California Berkeley
Haiyan Huang: University of California Berkeley
Amy E. Herr: University of California Berkeley

Nature Communications, 2021, vol. 12, issue 1, 1-14

Abstract: Abstract Multimeric cytoskeletal protein complexes orchestrate normal cellular function. However, protein-complex distributions in stressed, heterogeneous cell populations remain unknown. Cell staining and proximity-based methods have limited selectivity and/or sensitivity for endogenous multimeric protein-complex quantification from single cells. We introduce micro-arrayed, differential detergent fractionation to simultaneously detect protein complexes in hundreds of individual cells. Fractionation occurs by 60 s size-exclusion electrophoresis with protein complex-stabilizing buffer that minimizes depolymerization. Proteins are measured with a ~5-hour immunoassay. Co-detection of cytoskeletal protein complexes in U2OS cells treated with filamentous actin (F-actin) destabilizing Latrunculin A detects a unique subpopulation (~2%) exhibiting downregulated F-actin, but upregulated microtubules. Thus, some cells may upregulate other cytoskeletal complexes to counteract the stress of Latrunculin A treatment. We also sought to understand the effect of non-chemical stress on cellular heterogeneity of F-actin. We find heat shock may dysregulate filamentous and globular actin correlation. In this work, our assay overcomes selectivity limitations to biochemically quantify single-cell protein complexes perturbed with diverse stimuli.

Date: 2021
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-25212-3

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DOI: 10.1038/s41467-021-25212-3

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