Hydrogen-deuterium exchange studies on guanidinated pig heart lactate dehydrogenase

Abstract

Pig heart lactate dehydrogenase becomes more thermostable on increasing the degree of guanidination (conversion of lysine to homoarginine) (Minotani, N., Sekiguchi, T., Bautista, J.G. and Nosoh, Y. (1979) Biochim. Biophys. Acta 581, 334–341). The conformational change of the protein on guanidination was then examined by hydrogen-deuterium (H- 2H) exchange reactions. It was found that (i) the fluctuation degrees of peptides and tyrosine and tryptophan residues in the protein decrease in that order, (ii) two H- 2H exchangable tryptophan residues per subunit are freely accessible to solvent and the fluctuation degree of the residues does not change on guanidination, (iii) the H- 2H exchange detectable tyrosine residues are not freely accessible to solvent and become less fluctuating when 15 lysine residues per subunit are guanidinated, and (iv) the peptides become much less fluctuating on increasing the degree of guanidination. The specific activity of the enzyme decreased on guanidination. The increased thermostability of the protein on guanidination may be related to the decrease in flexibility of the molecular structure by sacrificing the enzyme activity.