5WOT

NMR solution structure of a-lytic protease using two 4D-spectra

Summary for 5WOT

• Entry DOI: 10.2210/pdb5wot/pdb
• NMR Information: BMRB: 30322
• Descriptor: Alpha-lytic protease (1 entity in total)
• Functional Keywords: protease, hydrolase
• Biological source: Lysobacter enzymogenes
• Total number of polymer chains: 1
• Total formula weight: 19875.13

Authors

Evangelidis, T.,Nerli, S.,Sgourakis, N.G.,Tripsianes, K. (deposition date: 2017-08-03, release date: 2018-02-07, Last modification date: 2024-10-23)

Primary citation

Evangelidis, T.,Nerli, S.,Novacek, J.,Brereton, A.E.,Karplus, P.A.,Dotas, R.R.,Venditti, V.,Sgourakis, N.G.,Tripsianes, K.
Automated NMR resonance assignments and structure determination using a minimal set of 4D spectra.
Nat Commun, 9:384-384, 2018

PubMed Abstract: Automated methods for NMR structure determination of proteins are continuously becoming more robust. However, current methods addressing larger, more complex targets rely on analyzing 6-10 complementary spectra, suggesting the need for alternative approaches. Here, we describe 4D-CHAINS/autoNOE-Rosetta, a complete pipeline for NOE-driven structure determination of medium- to larger-sized proteins. The 4D-CHAINS algorithm analyzes two 4D spectra recorded using a single, fully protonated protein sample in an iterative ansatz where common NOEs between different spin systems supplement conventional through-bond connectivities to establish assignments of sidechain and backbone resonances at high levels of completeness and with a minimum error rate. The 4D-CHAINS assignments are then used to guide automated assignment of long-range NOEs and structure refinement in autoNOE-Rosetta. Our results on four targets ranging in size from 15.5 to 27.3 kDa illustrate that the structures of proteins can be determined accurately and in an unsupervised manner in a matter of days.

PubMed: 29374165
DOI: 10.1038/s41467-017-02592-z

Experimental method

SOLUTION NMR