Yun-Sang Yu1, Ju-Young Lee2, Ji-Won Woo3, Jin-Se Kim1, Yoon-Hwan Bae4, Sung-Jin Cho5, Soon Cheol Park6. 1. School of Biological Sciences, Chungbuk National University, Cheongju, Chungbuk, 362-763, Republic of Korea. 2. Department of Biomedical Engineering, College of Medical Convergence, Catholic Kwandong University, Gangneung, Republic of Korea. 3. Department of Life Sciences, Chung-Ang University, 221 Heukseok-dong, Dongjak-gu, Seoul, 156-756, Republic of Korea. 4. Division of Life Sciences & Chemistry, Daejin University, Pocheon, Republic of Korea. 5. School of Biological Sciences, Chungbuk National University, Cheongju, Chungbuk, 362-763, Republic of Korea. sjchobio@cbnu.ac.kr. 6. Department of Life Sciences, Chung-Ang University, 221 Heukseok-dong, Dongjak-gu, Seoul, 156-756, Republic of Korea. scpark@cau.ac.kr.
Abstract
BACKGROUND: The invertebrate type (i-type) lysozyme not showing a clear homology with the known types of lysozyme was first demonstrated from a marine bivalve, conch and earthworm by N-terminal sequence. An i-type lysozyme isolated from the earthworm found to be up-regulated upon bacterial challenge, suggesting this lysozyme to function as an inducible immune factor. However, information on the i-type lysozyme related with digestive function is very limited in the earthworm. OBJECTIVE: The objective of this study is to investigate the molecular characteristics and function of the new i-type lysozyme from the earthworm. METHODS: To identify a new i-type lysozyme, multiple amino acid sequence alignment and phylogenetic analyses were employed. Its mRNA expression pattern was observed by fluorescent in situ hybridization (FISH). RESULTS: A new i-type lysozyme (Ea-iLys) from an earthworm, Eisenia andrei with the open reading frame of 678 bp (226 amino acid residues) appeared to comprise conserved 14 cysteine residues for disulfide bridges and amino acid residues for the enzyme activities of lysozyme and isopeptidase, of which mRNA expression is mainly localized in the lining of midgut epithelium. No significant expression signal was detected in immune competent sites such as chloragogue tissue, typhlosole region, body coelom and muscle layers. CONCLUSION: Our results suggest that this enzyme primarily acts as a digestive enzyme rather than an innate immune factor.
BACKGROUND: The invertebrate type (i-type) lysozyme not showing a clear homology with the known types of lysozyme was first demonstrated from a marine bivalve, conch and earthworm by N-terminal sequence. An i-type lysozyme isolated from the earthworm found to be up-regulated upon bacterial challenge, suggesting this lysozyme to function as an inducible immune factor. However, information on the i-type lysozyme related with digestive function is very limited in the earthworm. OBJECTIVE: The objective of this study is to investigate the molecular characteristics and function of the new i-type lysozyme from the earthworm. METHODS: To identify a new i-type lysozyme, multiple amino acid sequence alignment and phylogenetic analyses were employed. Its mRNA expression pattern was observed by fluorescent in situ hybridization (FISH). RESULTS: A new i-type lysozyme (Ea-iLys) from an earthworm, Eisenia andrei with the open reading frame of 678 bp (226 amino acid residues) appeared to comprise conserved 14 cysteine residues for disulfide bridges and amino acid residues for the enzyme activities of lysozyme and isopeptidase, of which mRNA expression is mainly localized in the lining of midgut epithelium. No significant expression signal was detected in immune competent sites such as chloragogue tissue, typhlosole region, body coelom and muscle layers. CONCLUSION: Our results suggest that this enzyme primarily acts as a digestive enzyme rather than an innate immune factor.