Abstract
One cDNA clone was purified from an adult Clonorchis sinensis cDNA library, and its deduced polypeptide sequence was found to be homologous with myosin regulatory light chain (MRLC) of invertebrates and vertebrates. Two amino-acid residues, Thr and Ser, were conserved at the phosphorylation sites that regulate the function of MRLCs. Recombinant C. sinensis MRLC (rCsMRLC) protein was produced and purified from Escherichia coli, and mouse anti-CsMRLC immune sera recognized a protein of molecular weight 24 kDa from a soluble protein preparation of C. sinensis. The CsMRLC protein was immunohistochemically localized to the muscle fibers of the subtegumental muscle layer and to the muscles of oral and ventral suckers. However, the rCsMRLC protein proved to be less useful antigen for the serodiagnosis of human clonorchiasis.
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References
Bruce AJ, Baumann BD, Hambly KH, Fajer PG (2001) The regulatory domain of the myosin head behaves as a rigid lever. Biochemistry 40:7868–7873
Choi D, Hong ST, Li S, Chung BS, Lim JH, Lee SH (2004) Bile duct changes in rats reinfected with Clonorchis sinensis. Korean J Parasitol 42:7–17
Devlin TM (1997) Textbook of biochemistry with clinical correlations, 4th edn. Wiley, New York, pp 946–959
Flavell DJ (1981) Liver-fluke infection as an aetiological factor in bile-duct carcinoma of man. Trans R Soc Trop Med Hyg 75:814–824
Hong SJ, Seong KY, Sohn WM, Song KY (2000) Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis. Mol Biochem Parasitol 108:207–216
Hong SJ, Kim TY, Gan XX, Sukontason K, Sukontason K, Kang SY (2002) Clonorchis sinensis: glutathione S-transferase as a serodiagnostic antigen for detecting IgG and IgE antibodies. Exp Parasitol 101:231–233
Hong SJ, Shin JK, Kang SY, Ryu JR (2003) Ultrastructural localization of phosphoglycerate kinase in adult Clonorchis sinensis. Parasitol Res 90:369–371
Ikebe M, Hartshorne DJ (1985a) Phosphorylation of smooth muscle myosin at two distinct sites by myosin light chain kinase. J Biol Chem 260:10027–10031
Ikebe M, Hartshorne DJ (1985b) Effects of Ca2+ on the conformation and enzymatic activity of smooth muscle myosin. J Biol Chem 260:13146–13153
Ikebe M, Hartshorne DJ, Elzinga M (1987) Identification, phosphorylation, and dephosphorylation of a second site for myosin light chain kinase on the 20,000-dalton light chain of smooth muscle myosin. J Biol Chem 261:36–39
Ikebe M, Ikebe R, Kamisoyama H, Reardon S, Schwonek JP, Sanders CR, Matsuura M (1994) Function of NH2-terminal domain of the regulatory light chain on the regulation of smooth muscle myosin. J Biol Chem 269:8173–8180
Ikebe M, Taketoshi K, Walter FS, Masataka S, Eisaku K (1998) A hinge at the central helix of the regulatory light chain of myosin is critical for phosphorylation-dependent regulation of smooth muscle myosin motor activity. J Biol Chem 273:17702–17707
Kang SY, Ahn IY, Park CY, Chung YB, Hong ST, Kong Y, Cho SY, Hong SJ (2001) Clonorchis sinensis: molecular cloning and characterization of 28-kDa glutathione S-transferase. Exp Parasitol 97:186–195
Kim TY, Kang SY, Park SH, Sukontason K, Sukontason K, Hong SJ (2001) Cystain capture enzyme-linked immunosorbent assay for serodiagnosis of human clonorchiasis and profile of captured antigenic protein of Clonorchis sinensis. Clin Diagn Lab Immunol 8:1076–1080
Lee JH, Yang HM, Bak UB, Rim HJ (1994) Promoting role of Clonorchis sinensis infection on induction of cholangiocarcinoma during two-step carcinogenesis. Korean J Parasitol 32:13–18
Lee JY, Kim TY, Gan XX, Kang SY, Hong SJ (2003) Use of a recombinant Clonorchis sinensis pore-forming peptide, clonorin, for serodiagnosis of clonorchiasis. Parasitol Res 52:175–178
Marston SB, Redwood CS (1991) The molecular anatomy of caldesmon. Biochem J 279:1–16
Onishi H, Umeda J, Uchiwa H, Watanabe S (1982) Purification of gizzard myosin light-chain phosphatase, reversible changes in the ATPase and superprecipitation activities of actomyosin in the presence of purified preparation of myosin light-chain phosphatase and kinase. J Biochem 91:265–271
Rim HJ (1986) The current pathobiology and chemotherapy of clonorchiasis. Korean J Parasitol 24(Suppl):7–69
Roger EK, Chang XJ, Kevin AE, Kulkarni S, Aguilera I, Kiehart DP (1991) The regulatory light chain of non-muscle myosin is encoded by spaghetti-squash, a gene required for cytokinesis in drosophila. Cell 65:1177–1189
Shin HR, Lee CU, Park HJ, Seol SY, Chung JM, Choi HC, Ahn YO, Shigemastu T (1996) Hepatitis B and C virus, Clonorchis sinensis for the risk of liver cancer: a case-control study in Pusan, Korea. Int J Epidemiol 25:933–940
Stull JT, Kamm KE, Krueger JK, Lin PJ, Katherine LP, Zhi G (1997) Ca2+ /calmodulin-dependent myosin light chain kinases. Signal Transduct Health Dis 31:141–150
Trybus K, Warshaw D (1991) In vitro evidence for smooth muscle crossbridge mechanical interactions. Adv Exp Med Biol 304:53–59
Uchimura T, Katsumi F, Yamamoto Y, Ueda K, Hosoya H (2002) Spatial localization of mono-and diphosphorylated myosin II regulatory light chain at the leading edge of motile HeLa cells. Cell Struct Funct 27:479–486
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This study was supported by a grant (01-PJ1-PG3-20200-0031) from Ministry of Health and Welfare, Republic of Korea.
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The nucleotide sequence reported herein was submitted to GenBank and assigned accession number AY519356.
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Kwon, YD., Cho, P.Y. & Hong, SJ. Clonorchis sinensis: molecular cloning and localization of myosin regulatory light chain. Parasitol Res 97, 21–26 (2005). https://doi.org/10.1007/s00436-005-1376-9
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DOI: https://doi.org/10.1007/s00436-005-1376-9