Smoothing membrane protein structure determination by initial upstream stage improvements
authors Pedro, AQ; Queiroz, JA; Passarinha, LA
nationality International
journal APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
author keywords Production; Membrane protein; Host; Codon usage; Protein 3D-structure; Structure determination; Quality control; Optimization
keywords ESCHERICHIA-COLI STRAINS; PICHIA-PASTORIS; MAMMALIAN-CELLS; CODON USAGE; RECOMBINANT EXPRESSION; FUNCTIONAL EXPRESSION; COUPLED RECEPTORS; N-GLYCOSYLATION; OUTER-MEMBRANE; E. COLI
abstract Membrane proteins (MP) constitute 20-30% of all proteins encoded by the genome of various organisms and perform a wide range of essential biological functions. However, despite they represent the largest class of protein drug targets, a relatively small number high-resolution 3D structures have been obtained yet. Membrane protein biogenesis is more complex than that of the soluble proteins and its recombinant biosynthesis has been a major drawback, thus delaying their further structural characterization. Indeed, the major limitation in structure determination of MP is the low yield achieved in recombinant expression, usually coupled to low functionality, pinpointing the optimization target in recombinant MP research. Recently, the growing attention that have been dedicated to the upstream stage of MP bioprocesses allowed great advances, permitting the evolution of the number of MP solved structures. In this review, we analyse and discuss effective solutions and technical advances at the level of the upstream stage using prokaryotic and eukaryotic organisms foreseeing an increase in expression yields of correctly folded MP and that may facilitate the determination of their three-dimensional structure. A section on techniques used to protein quality control and further structure determination of MP is also included. Lastly, a critical assessment of major factors contributing for a good decision-making process related to the upstream stage of MP is presented.
publisher SPRINGER
issn 0175-7598
year published 2019
volume 103
issue 14
beginning page 5483
ending page 5500
digital object identifier (doi) 10.1007/s00253-019-09873-1
web of science category Biotechnology & Applied Microbiology
subject category Biotechnology & Applied Microbiology
unique article identifier WOS:000473129900002
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journal impact factor 3.340
5 year journal impact factor 3.602
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