Patterns in the quinary structures of proteins. Plasticity and inequivalence of individual molecules in helical arrays of sickle cell hemoglobin and tubulin

Edelstein, Stuart J.

doi:10.1016/s0006-3495(80)84961-7

Cited by 37 publications

(23 citation statements)

References 37 publications

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“…Consequently, they have been termed quinary structures (Chien and Gierasch, 2014; Edelstein, 1980; Wallace et al, 2015). Formation of quinary structures often involves multivalent interactions between groups of binding partners (Li et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned Response

Riback

Katanski

Kear-Scott

et al. 2017

Cell

782

819

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Summary In eukaryotic cells, diverse stresses trigger coalescence of RNA-binding proteins into stress granules. In vitro, stress-granule-associated proteins can demix to form liquids, hydrogels, and other assemblies lacking fixed stoichiometry. Observing these phenomena has generally required conditions far removed from physiological stresses. We show that poly(A)-binding protein (Pab1 in yeast), a defining marker of stress granules, phase-separates and forms hydrogels in vitro upon exposure to physiological stress conditions. Other RNA-binding proteins depend upon low-complexity regions (LCRs) or RNA for phase separation, whereas Pab1’s LCR is not required for demixing, and RNA inhibits it. Based on unique evolutionary patterns, we create LCR mutations which systematically tune its biophysical properties and Pab1 phase separation in vitro and in vivo. Mutations which impede phase separation reduce organism fitness during prolonged stress. Poly(A)-binding protein thus acts as a physiological stress sensor, exploiting phase separation to precisely mark stress onset, a broadly generalizable mechanism.

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Section: Introductionmentioning

confidence: 99%

Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned Response

Riback

Katanski

Kear-Scott

et al. 2017

Cell

782

819

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“…Detailed characterization and quantification of the Caenorhabditis elegans proteome along its lifespan showed extensive proteome remodeling and suggested the sequestration of proteins in insoluble aggregates to be a protective strategy directed toward maintaining proteome integrity during aging [ 27 ]. In fact, the term quinary structures has been coined to describe such functional (stress-induced) large protein assemblies, as they lack the fixed stoichiometry specific to quaternary assemblies [ 28 , 29 ]. However, direct phenotypic evidence of their beneficial impact has remained limited.…”

Section: Introductionmentioning

confidence: 99%

Protein aggregates encode epigenetic memory of stressful encounters in individual Escherichia coli cells

et al. 2018

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Protein misfolding and aggregation are typically perceived as inevitable and detrimental processes tied to a stress- or age-associated decline in cellular proteostasis. A careful reassessment of this paradigm in the E. coli model bacterium revealed that the emergence of intracellular protein aggregates (PAs) was not related to cellular aging but closely linked to sublethal proteotoxic stresses such as exposure to heat, peroxide, and the antibiotic streptomycin. After removal of the proteotoxic stress and resumption of cellular proliferation, the polarly deposited PA was subjected to limited disaggregation and therefore became asymmetrically inherited for a large number of generations. Many generations after the original PA-inducing stress, the cells inheriting this ancestral PA displayed a significantly increased heat resistance compared to their isogenic, PA-free siblings. This PA-mediated inheritance of heat resistance could be reproduced with a conditionally expressed, intracellular PA consisting of an inert, aggregation-prone mutant protein, validating the role of PAs in increasing resistance and indicating that the resistance-conferring mechanism does not depend on the origin of the PA. Moreover, PAs were found to confer robustness to other proteotoxic stresses, as imposed by reactive oxygen species or streptomycin exposure, suggesting a broad protective effect. Our findings therefore reveal the potential of intracellular PAs to serve as long-term epigenetically inheritable and functional memory elements, physically referring to a previous cellular insult that occurred many generations ago and meanwhile improving robustness to a subsequent proteotoxic stress. The latter is presumably accomplished through the PA-mediated asymmetric inheritance of protein quality control components leading to their specific enrichment in PA-bearing cells.

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“…Here, the use of “quinary structure” is re-instituted to describe proteins having quaternary structure which aggregate. The quinary structure term was first proposed by Edelstein in 1980 in defining oligomeric hemoglobin structures causing sickle cell disease [3,4]. For monoclonal antibodies, quinary structure most correctly describes mAb oligomerization and is an important attribute of mAb therapeutics.…”

Section: Introductionmentioning

confidence: 99%

Simple NMR methods for evaluating higher order structures of monoclonal antibody therapeutics with quinary structure

Kang

Long

Lute

et al. 2016

Journal of Pharmaceutical and Biomedical Analysis

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Monoclonal antibody (mAb) drugs constitute the largest class of protein therapeutics currently on the market. Correctly folded protein higher order structure (HOS), including quinary structure, is crucial for mAb drug quality. The quinary structure is defined as the association of quaternary structures (e.g., oligomerized mAb). Here, several commonly available analytical methods, i.e., size-exclusion-chromatography (SEC) FPLC, multi-angle light scattering (MALS), circular dichroism (CD), NMR and multivariate analysis, were combined and modified to yield a complete profile of HOS and comparable metrics. Rituximab and infliximab were chosen for method evaluation because both IgG1 molecules are known to be homologous in sequence, superimposable in Fab crystal structure and identical in Fc structure. However, herein the two are identified to be significantly different in quinary structure in addition to minor secondary structure differences. All data collectively showed rituximab was mostly monomeric while infliximab was in mono-oligomer equilibrium driven by its Fab fragment. The quinary structure differences were qualitatively inferred from the less used but more reproducible dilution-injection-SEC-FPLC curve method. Quantitative principal component analysis (PCA) was performed on NMR spectra of either the intact or the in-situ enzymatic-digested mAb samples. The cleavage reactions happened directly in NMR tubes without further separation, which greatly enhanced NMR spectra quality and resulted in larger inter- and intra-lot variations based on PCA. The new in-situ enzymatic digestion method holds potential in identifying structural differences on larger therapeutic molecules using NMR.

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Patterns in the quinary structures of proteins. Plasticity and inequivalence of individual molecules in helical arrays of sickle cell hemoglobin and tubulin

Cited by 37 publications

References 37 publications

Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned Response

Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned Response

Protein aggregates encode epigenetic memory of stressful encounters in individual Escherichia coli cells

Simple NMR methods for evaluating higher order structures of monoclonal antibody therapeutics with quinary structure

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