Primary structure and function of novel O-glycosylated hirudins from the leech Hirudinaria manillensis

• Published in: Biochemistry (Easton) Vol. 31; no. 8; pp. 2294 - 2298
• Main Authors: Steiner, Verena, Knecht, Rene, Boernsen, K. Olaf, Gassmann, Ernst, Stone, Stuart R, Raschdorf, Fritz, Schlaeppi, Jean Marc, Maschler, Reinhard
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 01.03.1992
• Subjects: Amino Acid Sequence; Amino Acids - analysis; Aminoacids, peptides. Hormones. Neuropeptides; Analytical, structural and metabolic biochemistry; animal physiology; Animals; anticoagulants; biochemical composition; Biological and medical sciences; Carbohydrate Conformation; Freshwater; Fundamental and applied biological sciences. Psychology; Glycosylation; hirudin; Hirudinaria manillensis; Hirudins - chemistry; Hirudins - genetics; Hirudins - pharmacology; Leeches - chemistry; Molecular Sequence Data; molecular structure; Proteins; Sequence Alignment; Sequence Homology, Nucleic Acid; Structure-Activity Relationship; Thrombin - antagonists & inhibitors; variants; Composition; Molecular form; Peptides; Homology; Primary structure; Anticoagulant; Glycosylation; Annelida; Hirudo medicinalis; Hirudinea; Inhibition; Invertebrata; Aminoacid sequence
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00123a012

Hirudin from the leech Hirudo medicinalis is a most powerful anticoagulant, and many isoforms have been described. In the present work, the primary structure of two hirudins from the leech Hirudinaria manillensis has been elucidated. The antithrombotic activity is similar to that of H. medicinalis hirudins although the sequence identity is below 60%. Surprisingly, the hirudins were found to be glycosylated at one site. Sugar analysis after methanolysis yielded fucose, galactose, and N-acetylgalactosamine. These results combined with data from matrix-assisted laser desorption ionization mass spectrometry, plasma desorption mass spectrometry, capillary zone electrophoresis, and lectin-binding tests indicate that the sequence is Fuc-Gal beta 1-3GalNAc-(O-threonine). This structure shows an interesting similarity to human blood group H determinants.