Prediction of the determinants of thermal stability by linear discriminant analysis: the case of the glutamate dehydrogenase protein family.
Little is known about the determinants of thermal stability in individual protein families. Most of the knowledge on thermostability comes, in fact, from comparative analyses between large, and heterogeneous, sets of thermo- and mesophilic proteins. Here, we present a multivariate statistical approach aimed to detect signature sequences for thermostability in a single protein family. It was applied to the glutamate dehydrogenase (GDH) family, which is a good model for investigating this peculiar process. The structure of GDH consists of six subunits, each of them organized into two domains. Formation of ion-pair networks on the surface of the protein subunits, or increase in the inter-subunit hydrophobic interactions, have been suggested as important factors for explaining stability at high temperatures. However, identification of the amino acid changes that are involved in this process still remains elusive. Our approach consisted of a linear discriminant analysis on a set of GDH sequences from Archaea and Bacteria (33 thermo- and 36 mesophilic GDHs). It led to detection of 3 amino acid clusters as the putative determinants of thermal stability. They were localized at the subunit interface or in close proximity to the binding site of the NAD(P)(+) coenzyme. Analysis within the clusters led to prediction of 8 critical amino acid sites. This approach could have a wide utility, in the ligth of the notion that each protein family seems to adopt its own strategy for achieving thermostability.
Department of Life Sciences, University of Parma, Parco Area delle Scienze 11/A, I-43124 Parma, Italy. Electronic address: angelo.pavesi@unipr.it.
Angelo
A
Pavesi
D019843:Archaeal Proteins / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D004795:Enzyme Stability; D005969:Glutamate Dehydrogenase / Q000737:chemistry*; D006358:Hot Temperature; D008956:Models, Chemical; D021122:Protein Subunits / Q000737:chemistry
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D021122:Protein Subunits; D005969:Glutamate Dehydrogenase
Archaea; Bacteria; Mesophile; Multivariate statistics; Thermophile