Prp40 Homolog A Is a Novel Centrin Target.

Díaz Casas A, Chazin WJ, Pastrana-Ríos B
Biophys J. 2017 112 (12): 2529-2539

PMID: 28636910 · PMCID: PMC5479147 · DOI:10.1016/j.bpj.2017.03.042

Pre-mRNA processing protein 40 (Prp40) is a nuclear protein that has a role in pre-mRNA splicing. Prp40 possesses two leucine-rich nuclear export signals, but little is known about the function of Prp40 in the export process. Another protein that has a role in protein export is centrin, a member of the EF-hand superfamily of Ca-binding proteins. Prp40 was found to be a centrin target by yeast-two-hybrid screening using both Homo sapiens centrin 2 (Hscen2) and Chlamydomonas reinhardtii centrin (Crcen). We identified a centrin-binding site within H. sapiens Prp40 homolog A (HsPrp40A), which contains a hydrophobic triad WLL that is known to be important in the interaction with centrin. This centrin-binding site is highly conserved within the first nuclear export signal consensus sequence identified in Saccharomyces cerevisiae Prp40. Here, we examine the interaction of HsPrp40A peptide (HsPrp40Ap) with both Hscen2 and Crcen by isothermal titration calorimetry. We employed the thermodynamic parameterization to estimate the polar and apolar surface area of the interface. In addition, we have defined the molecular mechanism of thermally induced unfolding and dissociation of the Crcen-HsPrp40Ap complex using two-dimensional infrared correlation spectroscopy. These complementary techniques showed for the first time, to our knowledge, that HsPrp40Ap interacts with centrin in vitro, supporting a coupled functional role for these proteins in pre-mRNA splicing.

Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

MeSH Terms (14)

Binding Sites Calorimetry Carrier Proteins Chlamydomonas reinhardtii Circular Dichroism Humans Hydrophobic and Hydrophilic Interactions Protein Unfolding Recombinant Proteins Sequence Homology, Amino Acid Spectroscopy, Fourier Transform Infrared Thermodynamics Trimethoprim, Sulfamethoxazole Drug Combination Two-Hybrid System Techniques

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