DOI: 10.1101/498287Dec 16, 2018Paper

Integrated NMR and cryo-EM atomic-resolution structure determination of a half-megadalton enzyme complex

BioRxiv : the Preprint Server for Biology
Diego F GautoPaul Schanda

Abstract

Atomic-resolution structure determination is the key requirement for understanding protein function. Cryo-EM and NMR spectroscopy both provide structural information, but currently cryo-EM does not routinely give access to atomic-level structural data, and, generally, NMR structure determination is restricted to small (<30 kDa) proteins. We introduce an integrated structure determination approach that simultaneously uses NMR and EM data to overcome the limits of each of these methods. The approach enabled determination of the high-resolution structure of the 468 kDa large dodecameric aminopeptidase TET2 to a precision and accuracy below 1 Angstrom by combining secondary-structure information obtained from near-complete magic-angle-spinning NMR assignments of the 39 kDa-large subunits, distance restraints from backbone amides and specifically labelled methyl groups, and a 4.1 Angstrom resolution EM map. The resulting structure exceeds current standards of NMR and EM structure determination in terms of molecular weight and precision. Importantly, the approach is successful even in cases where only medium-resolution (up to 8 Angstrom) cryo-EM data are available, thus paving avenues for the structure determination of challenging bi...Continue Reading

Related Concepts

Electron Spin Resonance Spectroscopy
Electron Microscopy
Comprehension
Protein Structure, Secondary
Resolut
Structure
Methylate
Spectroscopy, Nuclear Magnetic Resonance
Determination Aspects
Probable Methylcytosine Dioxygenase TET2

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