Efficient protein production inspired by how spiders make silk

Membrane proteins are targets of most available pharmaceuticals, but they are difficult to produce recombinantly, like many other aggregation-prone proteins. Spiders can produce silk proteins at huge concentrations by sequestering their aggregation-prone regions in micellar structures, where the ver...

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Veröffentlicht in:	Nature communications Jg. 8; H. 1; S. 15504 - 15
Hauptverfasser:	Kronqvist, Nina, Sarr, Médoune, Lindqvist, Anton, Nordling, Kerstin, Otikovs, Martins, Venturi, Luca, Pioselli, Barbara, Purhonen, Pasi, Landreh, Michael, Biverstål, Henrik, Toleikis, Zigmantas, Sjöberg, Lisa, Robinson, Carol V., Pelizzi, Nicola, Jörnvall, Hans, Hebert, Hans, Jaudzems, Kristaps, Curstedt, Tore, Rising, Anna, Johansson, Jan
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	London Nature Publishing Group UK 23.05.2017 Nature Publishing Group Nature Portfolio
Schlagworte:	101/28 140/131 631/1647/2230/2233 631/337/470/2284 631/61/338/469 82/16 82/29 82/58 82/80 82/83 Amino acids Animal diseases Animal models Animals Biochemistry Cholecystokinin - chemistry Chromatography Circular Dichroism Dimerization Disease Models, Animal E coli Escherichia coli - metabolism Female Fibroins - biosynthesis Humanities and Social Sciences Hydrogen-Ion Concentration Lung - pathology Magnetic Resonance Spectroscopy Medical Bioscience Medicinsk biovetenskap Micelles Microscopy, Electron, Transmission multidisciplinary Mutagenesis, Site-Directed Mutation Peptides Peptides - chemistry Protein Domains Proteins Rabbits Recombinant Proteins - biosynthesis Respiration Disorders - drug therapy Respiratory diseases Science Science (multidisciplinary) Silk Silk - biosynthesis Spiders Surface-Active Agents - chemistry Surfactants Sweden
ISSN:	2041-1723, 2041-1723
Online-Zugang:	Volltext
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Zusammenfassung:	Membrane proteins are targets of most available pharmaceuticals, but they are difficult to produce recombinantly, like many other aggregation-prone proteins. Spiders can produce silk proteins at huge concentrations by sequestering their aggregation-prone regions in micellar structures, where the very soluble N-terminal domain (NT) forms the shell. We hypothesize that fusion to NT could similarly solubilize non-spidroin proteins, and design a charge-reversed mutant (NT) that is pH insensitive, stabilized and hypersoluble compared to wild-type NT. NT-transmembrane protein fusions yield up to eight times more of soluble protein in Escherichia coli than fusions with several conventional tags. NT* enables transmembrane peptide purification to homogeneity without chromatography and manufacture of low-cost synthetic lung surfactant that works in an animal model of respiratory disease. NT* also allows efficient expression and purification of non-transmembrane proteins, which are otherwise refractory to recombinant production, and offers a new tool for reluctant proteins in general. The properties of many transmembrane or aggregation-prone proteins make them difficult to recombinantly express. Here the authors use a modified N-terminal domain of a spider silk protein to express and purify several difficult to express proteins at levels considerably higher than with conventional tags.
Bibliographie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23
ISSN:	2041-1723 2041-1723
DOI:	10.1038/ncomms15504