X-ray crystallography and structural stability of digestive lysozyme from cow stomach

• Published in: The FEBS journal Vol. 276; no. 8; pp. 2192 - 2200
• Main Authors: Nonaka, Yasuhiro, Akieda, Daisuke, Aizawa, Tomoyasu, Watanabe, Nobuhisa, Kamiya, Masakatsu, Kumaki, Yasuhiro, Mizuguchi, Mineyuki, Kikukawa, Takashi, Demura, Makoto, Kawano, Keiichi
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01.04.2009; Blackwell Publishing Ltd
• Subjects: Animals; Biochemistry; Cattle; Crystallography; Crystallography, X-Ray; Enzymes; Gastroenterology; Guanidine - pharmacology; Hydrogen-Ion Concentration; lysozyme; Models, Molecular; molecular evolution; Muramidase - chemistry; Muramidase - metabolism; protease resistance; Protein Conformation; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Ruminantia; Stomach - enzymology; structural stability; Veterinary medicine; X-ray diffraction; X‐ray crystallography
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/j.1742-4658.2009.06948.x

In ruminants, some leaf-eating animals, and some insects, defensive lysozymes have been adapted to become digestive enzymes, in order to digest bacteria in the stomach. Digestive lysozyme has been reported to be resistant to protease and to have optimal activity at acidic pH. The structural basis of the adaptation providing persistence of lytic activity under severe gastric conditions remains unclear. In this investigation, we obtained the crystallographic structure of recombinant bovine stomach lysozyme 2 (BSL2). Our denaturant and thermal unfolding experiments revealed that BSL2 has high conformational stability at acidic pH. The high stability in acidic solution could be related to pepsin resistance, which has been previously reported for BSL2. The crystal structure of BSL2 suggested that negatively charged surfaces, a shortened loop and salt bridges could provide structural stability, and thus resistance to pepsin. It is likely that BSL2 loses lytic activity at neutral pH because of adaptations to resist pepsin.