| Naslov: | Droplet-based screening of phosphate transfer catalysis reveals how epistasis shapes MAP kinase interactions with substrates |
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| Avtorji: | ID Scheele, Remkes A. (Avtor)
ID Lindenburg, Laurens H. (Avtor)
ID Petek, Maya (Avtor)
ID Schober, Markus (Avtor)
ID Dalby, Kevin N. (Avtor)
ID Hollfelder, Florian (Avtor) |
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| Datoteke: |  Scheele-2022-Droplet-based_screening_of_phosph.pdf (2,42 MB)
MD5: A49086104F829195C0E5737E82F4F3E1
 https://doi.org/10.1038/s41467-022-28396-4
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| Jezik: | Angleški jezik |
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| Vrsta gradiva: | Znanstveno delo |
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| Tipologija: | 1.01 - Izvirni znanstveni članek |
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| Organizacija: | MF - Medicinska fakulteta
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| Opis: | The combination of ultrahigh-throughput screening and sequencing informs on function and intragenic epistasis within combinatorial protein mutant libraries. Establishing a droplet-based, in vitro compartmentalised approach for robust expression and screening of protein kinase cascades (>107 variants/day) allowed us to dissect the intrinsic molecular features of the MKK-ERK signalling pathway, without interference from endogenous cellular components. In a six-residue combinatorial library of the MKK1 docking domain, we identified 29,563 sequence permutations that allow MKK1 to efficiently phosphorylate and activate its downstream target kinase ERK2. A flexibly placed hydrophobic sequence motif emerges which is defined by higher order epistatic interactions between six residues, suggesting synergy that enables high connectivity in the sequence landscape. Through positive epistasis, MKK1 maintains function during mutagenesis, establishing the importance of co-dependent residues in mammalian protein kinase-substrate interactions, and creating a scenario for the evolution of diverse human signalling networks. |
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| Ključne besede: | protein, epistasis, catalysis, phosphate transfer, MAP kinase |
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| Status publikacije: | Objavljeno |
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| Verzija publikacije: | Objavljena publikacija |
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| Poslano v recenzijo: | 07.04.2021 |
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| Datum sprejetja članka: | 10.01.2022 |
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| Datum objave: | 11.02.2022 |
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| Založnik: | Nature Publishing Group |
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| Leto izida: | 2022 |
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| Št. strani: | Str. 1-14 |
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| Številčenje: | Letn. 13, Št. članka 844 |
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| PID: | 20.500.12556/DKUM-89347  |
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| UDK: | 577 |
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| COBISS.SI-ID: | 98894851 |
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| DOI: | 10.1038/s41467-022-28396-4 |
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| ISSN pri članku: | 2041-1723 |
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| Datum objave v DKUM: | 04.07.2024 |
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| Število ogledov: | 252 |
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| Število prenosov: | 20 |
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| Metapodatki: |    |
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| Področja: | Ostalo
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