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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Bibliographic Details
Published in:Nature communications Vol. 8; no. 1; pp. 15504 - 15
Main Authors: 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
Language:English
Published: London Nature Publishing Group UK 23.05.2017
Nature Publishing Group
Nature Portfolio
Subjects:
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 Access:Get full text
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Summary: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.
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ISSN:2041-1723
2041-1723
DOI:10.1038/ncomms15504