Maltose Binding Protein Is Partially Structured in Its Molten Globule State

Reichenwallner, Joerg and Chakour, Mohammed and Indu, S and Varadarajan, Raghavan and Trommer, Wolfgang E (2013) Maltose Binding Protein Is Partially Structured in Its Molten Globule State. In: APPLIED MAGNETIC RESONANCE, 44 (8). pp. 983-995.

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Abstract

Seven double cysteine mutants of maltose binding protein (MBP) were generated with one each in the active cleft at position 298 and the second cysteine distributed over both domains of the protein. These cysteines were spin labeled and distances between the labels in biradical pairs determined by pulsed double electron-electron resonance (DEER) measurements. The values were compared with theoretical predictions of distances between the labels in biradicals constructed by molecular modeling from the crystal structure of MBP without maltose and were found to be in excellent agreement. MBP is in a molten globule state at pH 3.3 and is known to still bind its substrate maltose. The nitroxide spin label was sufficiently stable under these conditions. In preliminary experiments, DEER measurements were carried out with one of the mutants yielding a broad distance distribution as was to be expected if there is no explicit tertiary structure and the individual helices pointing into all possible directions.

Item Type:	Journal Article
Publication:	APPLIED MAGNETIC RESONANCE
Publisher:	SPRINGER WIEN
Additional Information:	Copyright for this article belongs to Springer
Department/Centre:	Division of Mechanical Sciences > Chemical Engineering
Date Deposited:	08 Oct 2013 11:23
Last Modified:	08 Oct 2013 11:23
URI:	http://eprints.iisc.ac.in/id/eprint/47523

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