title: Larval and pupal induction and N-terminal amino acid sequence of lysozyme from Heliothis virescens.
abstract: Fifth instar larvae and prepupae of Heliothis virescens (tobacco budworm) were injected with live Enterobacter cloacae and bled at different times after vaccination. Immune pupal hemolymph showed a 54 times increase in lysozyme activity when compared with normal larval hemolymph, and an 11 times increase of lysozyme activity when compared with immune larval hemolymph. Lysozyme activity of the normal pupal hemolymph increased as greatly as did lysozyme activity of the immune larval hemolymph after metamorphosis. The pupal immune response with regard to lysozyme was much greater than the larval immune response in H. virescens. Lysozyme was purified by heat treatment at 100 degrees C and a chromatography series that included reverse-phase HPLC. The molecular mass of H. virescens lysozyme was approximately 16 kDa by SDS-PAGE which is greater than other insect lysozymes and chicken lysozyme. Amino acid sequence of the N-terminus showed that H. virescens lysozyme is 82% homologous with lysozyme of Manduca sexta and Galleria mellonella. CNBr cleavage of H. virescens lysozyme produced 11 and 6 kDa peptide fragments indicating that one methionine was present, which was also supported by amino acid analysis. However, methionine was located at the carboxyl terminal side rather than the N-terminal side as judged by the N-terminal sequences of each peptide fragment. The residue 22 in most lepidopteran lysozymes is methionine, whereas H. virescens lysozyme had a leucine at residue 22. There was an amino acid deletion near the carboxyl terminal side of H. virescens lysozyme as also found in Trichoplusia ni.
authors.affiliation: Department of Microbiology and Molecular Cell Sciences, University of Memphis, Memphis, TN, USA
authors.forename: K T.
authors.initials: KT
authors.lastname: Chung
authors.forename: D D.
authors.initials: DD
authors.lastname: Ourth