A systematic family-wide investigation reveals that ~30% of mammalian PDZ domains engage in PDZ-PDZ interactions

PDZ domains are independently folded modules that typically mediate protein-protein interactions by binding to the C termini of their target proteins. However, in a few instances, PDZ domains have been reported to dimerize with other PDZ domains. To investigate this noncanonical-binding mode further, we used protein microarrays comprising virtually every mouse PDZ domain to systematically query all possible PDZ-PDZ pairs. We then used fluorescence polarization to retest and quantify interactions and coaffinity purification to test biophysically validated interactions in the context of their full-length proteins. Overall, we discovered 37 PDZ-PDZ interactions involving 46 PDZ domains (~30% of all PDZ domains tested), revealing that dimerization is a more frequently used binding mode than was previously appreciated. This suggests that many PDZ domains evolved to form multiprotein complexes by simultaneously interacting with more than one ligand.     

Effect of poly (lactic acid)‐graft‐glycidyl methacrylate as a compatibilizer on properties of poly (lactic acid)/banana fiber biocomposites

The main aim of this study was to synthesis of poly (lactic acid) (PLA)‐graft‐glycidyl methacrylate (GMA) as well as its influence on the properties of PLA/banana fiber biocomposites. PLA‐graft‐GMA graft copolymer (GC) was synthesized by melt blending PLA with GMA using benzoyl peroxide and dicumyl peroxide as initiators. Graft copolymerization was confirmed by FTIR and ¹H‐NMR spectroscopic studies. PLA/silane treated banana fiber (SiB) biocomposites with various GC concentrations were prepared by melt blending followed by injection molding techniques. The influence of GC content on the mechanical, thermal and moisture resistance properties of the composite was investigated. The addition of 15 wt% GC content in the biocomposite provided optimum tensile and flexural strength, which is attributed to the greater compatibility between fiber and PLA matrix. The thermal properties of biocomposites have been evaluated using thermogravimetric analysis which provided evidence of improved interfacial adhesion between SiB and PLA by the addition of GC. Additionally, GC enhanced the moisture absorption resistance of biocomposites. These results indicated that GC is indeed a good candidate as a compatibilizing agent to improve the compatibility in PLA/fiber biocomposites. Copyright © 2015 John Wiley & Sons, Ltd.     

Uncovering quantitative protein interaction networks for mouse PDZ domains using protein microarrays

One of the principal challenges in systems biology is to uncover the networks of protein-protein interactions that underlie most biological processes. To date, experimental efforts directed at this problem have largely produced only qualitative networks that are replete with false positives and false negatives. Here, we describe a domain-centered approach--compatible with genome-wide investigations--that enables us to measure the equilibrium dissociation constant (K(D)) of recombinant PDZ domains for fluorescently labeled peptides that represent physiologically relevant binding partners. Using a pilot set of 22 PDZ domains, 4 PDZ domain clusters, and 20 peptides, we define a gold standard dataset by determining the K(D) for all 520 PDZ-peptide combinations using fluorescence polarization. We then show that microarrays of PDZ domains identify interactions of moderate to high affinity (K(D) < or = 10 microM) in a high-throughput format with a false positive rate of 14% and a false negative rate of 14%. By combining the throughput of protein microarrays with the fidelity of fluorescence polarization, our domain/peptide-based strategy yields a quantitative network that faithfully recapitulates 85% of previously reported interactions and uncovers new biophysical interactions, many of which occur between proteins that are co-expressed. From a broader perspective, the selectivity data produced by this effort reveal a strong concordance between protein sequence and protein function, supporting a model in which interaction networks evolve through small steps that do not involve dramatic rewiring of the network.