1. Structure, molecular evolution, and hydrolytic specificities of largemouth bass pepsins.
- Author
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Miura Y, Suzuki-Matsubara M, Kageyama T, and Moriyama A
- Subjects
- Amino Acid Sequence, Animals, Bass classification, Bass metabolism, Cloning, Molecular, DNA, Complementary metabolism, Escherichia coli genetics, Escherichia coli metabolism, Fish Proteins chemistry, Fish Proteins metabolism, Gene Expression, Hydrolysis, Hydrophobic and Hydrophilic Interactions, Models, Molecular, Molecular Sequence Data, Pepsin A chemistry, Pepsin A genetics, Pepsin A metabolism, Pepsinogens chemistry, Pepsinogens metabolism, Phylogeny, Protein Isoforms chemistry, Protein Isoforms genetics, Protein Isoforms metabolism, Protein Structure, Secondary, Protein Structure, Tertiary, RNA, Messenger genetics, RNA, Messenger metabolism, Sequence Alignment, Substrate Specificity, Swine, Thermodynamics, Bass genetics, DNA, Complementary genetics, Evolution, Molecular, Fish Proteins genetics, Pepsinogens genetics
- Abstract
The nucleotide sequences of largemouth bass pepsinogens (PG1, 2 and 3) were determined after molecular cloning of the respective cDNAs. Encoded PG1, 2 and 3 were classified as fish pepsinogens A1, A2 and C, respectively. Molecular evolutionary analyses show that vertebrate pepsinogens are classified into seven monophyletic groups, i.e. pepsinogens A, F, Y (prochymosins), C, B, and fish pepsinogens A and C. Regarding the primary structures, extensive deletion was obvious in S'1 loop residues in fish pepsin A as well as tetrapod pepsin Y. This deletion resulted in a decrease in hydrophobic residues in the S'1 site. Hydrolytic specificities of bass pepsins A1 and A2 were investigated with a pepsin substrate and its variants. Bass pepsins preferred both hydrophobic/aromatic residues and charged residues at the P'1 sites of substrates, showing the dual character of S'1 sites. Thermodynamic analyses of bass pepsin A2 showed that its activation Gibbs energy change (∆G(‡)) was lower than that of porcine pepsin A. Several sites of bass pepsin A2 moiety were found to be under positive selection, and most of them are located on the surface of the molecule, where they are involved in conformational flexibility. The broad S'1 specificity and flexible structure of bass pepsin A2 are thought to cause its high proteolytic activity., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2016
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