5NDA

NMR Structural Characterisation of Pharmaceutically Relevant Proteins Obtained Through a Novel Recombinant Production: The Case of The Pulmonary Surfactant Polypeptide C Analogue rSP-C33Leu.

5NDA の概要

• エントリーDOI: 10.2210/pdb5nda/pdb
• NMR情報: BMRB: 34114
• 分子名称: rSP-C33Leu -RECOMBINANT PULMONARY SURFACTANT-ASSOCIATED POLYPEPTIDE C ANALOGUE- (1 entity in total)
• 機能のキーワード: protein, pulmonary surfactant protein, sp-c analogue, recombinant protein, solubility tag, fusion protein, nt domain, protein structure
• 由来する生物種: Sus scrofa (Pig)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 3600.73

構造登録者

Venturi, L.,Pioselli, B.,Johansson, J.,Rising, A.,Kronqvist, N.,Nordling, K. (登録日: 2017-03-08, 公開日: 2017-06-07, 最終更新日: 2024-06-19)

主引用文献

Kronqvist, N.,Sarr, M.,Lindqvist, A.,Nordling, K.,Otikovs, M.,Venturi, L.,Pioselli, B.,Purhonen, P.,Landreh, M.,Biverstal, H.,Toleikis, Z.,Sjoberg, L.,Robinson, C.V.,Pelizzi, N.,Jornvall, H.,Hebert, H.,Jaudzems, K.,Curstedt, T.,Rising, A.,Johansson, J.
Efficient protein production inspired by how spiders make silk.
Nat Commun, 8:15504-15504, 2017

PubMed Abstract: 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.

PubMed: 28534479
DOI: 10.1038/ncomms15504

実験手法

SOLUTION NMR