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Protein Tyrosine Phosphatase Oligomerization Studied by a Combination of ¹⁵N NMR Relaxation and ¹²⁹Xe NMR. Effect of Buffer Containing Arginine and Glutamic Acid

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Jascha Blobel,^† Sabine Schmidl,^‡ David Vidal,^† Lydia Nisius,^‡ Pau Bernadó,^† Oscar Millet,^†^# Eike Brunner,^‡ and Miquel Pons*^†^§

Contribution from the Laboratory of Biomolecular NMR, Institute for Research in Biomedicine, Parc Cientfic de Barcelona, Josep Samitier, 1-5, E-08028 Barcelona, Spain, Institut fr Biophysik und Physikalische Biochemie, Universitt Regensburg, 93040 Regensburg, Germany, and Departament de Qumica Orgnica, Universitat de Barcelona, Mart i Franqus, 1-11, E-08028 Barcelona, Spain

J. Am. Chem. Soc., 2007, 129 (18), pp 5946–5953

DOI: 10.1021/ja069144p

Publication Date (Web): April 17, 2007

†

Institute for Research in Biomedicine.

‡

Universität Regensburg.

Present address: Structural Biology Unit, CIC bioGUNE, Bizkaia Technology Park, Building 801A, 48160 Derio, Spain.

In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the paper should be addressed.

Universitat de Barcelona.

, mpons@ub.edu

Abstract

¹⁵N NMR relaxation and ¹²⁹Xe NMR chemical shift measurements offer complementary information to study weak protein−protein interactions. They have been applied to study the oligomerization equilibrium of a low-molecular-weight protein tyrosine phosphatase in the presence of 50 mM arginine and 50 mM glutamic acid. These experimental conditions are shown to enhance specific protein−protein interactions while decreasing nonspecific aggregation. In addition, ¹²⁹Xe NMR chemical shifts become selective reporters of one particular oligomer in the presence of arginine and glutamic acid, indicating that a specific Xe binding site is created in the oligomerization process. It is suggested that the multiple effects of arginine and glutamic acid are related to their effective excluded volume that favors specific protein association and the destabilization of partially unfolded forms that preferentially interact with xenon and are responsible for nonspecific protein aggregation.

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Published In Issue May 09, 2007
Received December 29, 2006

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Article

Protein Tyrosine Phosphatase Oligomerization Studied by a Combination of ¹⁵N NMR Relaxation and ¹²⁹Xe NMR. Effect of Buffer Containing Arginine and Glutamic Acid