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Auto-inducing media for uniform isotope labeling of proteins with 15N, 13C and 2H.

Guthertz, Nicolas and Klopp, Julia and Winterhalter, Aurelie and Fernandez, Cesar and Gossert, Alvar (2015) Auto-inducing media for uniform isotope labeling of proteins with 15N, 13C and 2H. Journal of Biomolecular NMR, 62 (2). pp. 169-177.

Abstract

An auto-inducing medium for uniform isotope labelling of proteins with 15N, 13C and/or 2H is presented. So far, auto-inducing media have not found widespread application in the NMR field, because of the prohibitively high cost of labeled lactose, which is an essential ingredient of such media. Here, we propose using lactose that is only selectively labeled on the glucose moiety. It can be produced from inexpensive and readily available substrates: labeled glucose and unlabeled activated galactose. With this approach, uniformly isotope labeled proteins were produced in unattended auto-inducing cultures with incorporation of 13C, 15N of 96.6% and 2H,15N of 98.8%, as determined by mass spectroscopy. With the present protocol, the NMR community could profit from the many advantages that auto-inducing media offer.

Item Type: Article
Date Deposited: 14 Jun 2016 23:45
Last Modified: 04 Jul 2016 23:45
URI: https://oak.novartis.com/id/eprint/22247

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