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Production of unstable proteins through the formation of stable core complexes.

ABSTRACT: Purification of proteins that participate in large transient complexes is impeded by low amounts, heterogeneity, instability and poor solubility. To circumvent these difficulties we set up a methodology that enables the production of stable complexes for structural and functional studies. This procedure is benchmarked and applied to two challenging protein families: the human steroid nuclear receptors (SNR) and the HIV-1 pre-integration complex. In the context of transcriptional regulation studies, we produce and characterize the ligand-binding domains of the glucocorticoid nuclear receptor and the oestrogen receptor beta in complex with a TIF2 (transcriptional intermediary factor 2) domain containing the three SNR-binding motifs. In the context of retroviral integration, we demonstrate the stabilization of the HIV-1 integrase by formation of complexes with partner proteins and DNA. This procedure provides a powerful research tool for structural and functional studies of proteins participating in non-covalent macromolecular complexes.

SUBMITTER: Levy N

PROVIDER: S-EPMC4800440 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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Production of unstable proteins through the formation of stable core complexes.

Levy Nicolas N Eiler Sylvia S Pradeau-Aubreton Karine K Maillot Benoit B Stricher François F Ruff Marc M

Nature communications 20160317

Purification of proteins that participate in large transient complexes is impeded by low amounts, heterogeneity, instability and poor solubility. To circumvent these difficulties we set up a methodology that enables the production of stable complexes for structural and functional studies. This procedure is benchmarked and applied to two challenging protein families: the human steroid nuclear receptors (SNR) and the HIV-1 pre-integration complex. In the context of transcriptional regulation studi ...[more]

PMID: 26983699

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Project description:BackgroundIt has been demonstrated that core strength training is an effective means to enhance trunk muscle strength (TMS) and proxies of physical fitness in youth. Of note, cross-sectional studies revealed that the inclusion of unstable elements in core strengthening exercises produced increases in trunk muscle activity and thus provide potential extra training stimuli for performance enhancement. Thus, utilizing unstable surfaces during core strength training may even produce larger performance gains. However, the effects of core strength training using unstable surfaces are unresolved in youth. This randomized controlled study specifically investigated the effects of core strength training performed on stable surfaces (CSTS) compared to unstable surfaces (CSTU) on physical fitness in school-aged children.MethodsTwenty-seven (14 girls, 13 boys) healthy subjects (mean age: 14 ± 1 years, age range: 13-15 years) were randomly assigned to a CSTS (n = 13) or a CSTU (n = 14) group. Both training programs lasted 6 weeks (2 sessions/week) and included frontal, dorsal, and lateral core exercises. During CSTU, these exercises were conducted on unstable surfaces (e.g., TOGU© DYNAIR CUSSIONS, THERA-BAND© STABILITY TRAINER).ResultsSignificant main effects of Time (pre vs. post) were observed for the TMS tests (8-22%, f = 0.47-0.76), the jumping sideways test (4-5%, f = 1.07), and the Y balance test (2-3%, f = 0.46-0.49). Trends towards significance were found for the standing long jump test (1-3%, f = 0.39) and the stand-and-reach test (0-2%, f = 0.39). We could not detect any significant main effects of Group. Significant Time x Group interactions were detected for the stand-and-reach test in favour of the CSTU group (2%, f = 0.54).ConclusionsCore strength training resulted in significant increases in proxies of physical fitness in adolescents. However, CSTU as compared to CSTS had only limited additional effects (i.e., stand-and-reach test). Consequently, if the goal of training is to enhance physical fitness, then CSTU has limited advantages over CSTS.Trial registrationClinicalTrials.gov Identifier: NCT02290457 Registered 13 November 2014.

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Production of unstable proteins through the formation of stable core complexes.

Publications

Production of unstable proteins through the formation of stable core complexes.

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