Characterization of a complementary deoxyribonucleic acid for the coagulogen of Limulus polyphemus |
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| Affiliation: | 1. Department of Materials Science and Engineering, Stony Brook University, Stony Brook, NY 11794, USA;2. Department of Pathology, Stony Brook University, Stony Brook, NY 11794, USA;3. Food and Drug Administration, Center for Devices and Radiological Health, Silver Spring, MD 20903, USA;4. Department of Oral Biology and Pathology, School of Dental Medicine, Stony Brook University, Stony Brook, NY 11794, USA;1. Department of Chemical Engineering, Northeastern University, Boston, MA 02115-5000, USA;2. Biomaterials Innovations Research Center, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA;3. Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA;4. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA;5. Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX, USA;6. Department of Physics, King Abdulaziz University, Jeddah 21569, Saudi Arabia;1. Department of Rheumatology and Immunology, Institute of Translational Medicine, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, China;2. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China;1. Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, People’s Republic of China;2. College of Chemical Engineering, Fuzhou University, Fuzhou, 350108, People’s Republic of China;3. Beijing National Laboratory for Molecular Sciences, State Key Laboratory of Polymer Physics and Chemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, People’s Republic of China;4. Department of General Surgery, The Second Hospital of Jilin University, Changchun, 130041, People’s Republic of China;5. College of Chemistry, Fuzhou University, Fuzhou, 350116, People’s Republic of China;6. Jilin Biomedical Polymers Engineering Laboratory, Changchun, 130022, People’s Republic of China |
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| Abstract: | An 869-nucleotide-long cDNA clone for the coagulogen from Limulus amebocyte has been isolated and its nucleotide sequence has been determined. The deduced amino-acid sequence revealed a signal peptide, 20 amino acids long, and a mature protein of 175 amino acids. The amino-acid sequence of the coagulogen was compared to all known proteins by two computer programs. Using these programs, Limulus coagulogen showed 70% homology with the coagulogen of Tachypleus tridentatus (Japanese horseshoe crab). Further computer analysis showed no statistically significant homology to support an evolutionary origin of the horseshoe crab coagulogen common to other protein families. These results place horseshoe crab coagulogen in a new superfamily unrelated to any other proteins investigated. RNA blot analysis of Limulus RNA indicated that the coagulogen mRNA was about 900 bases long and represented an abundant species in the amebocyte while detected only in small quantities in the hepatopancreas. Besides mature RNA, high-molecular-weight forms of coagulogen RNA were also observed. Southern blot analysis of Limulus DNA digested with restriction endonucleases suggested that the Limulus coagulogen gene contains at least three introns, or belongs to a multigene family. |
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