Valenzuela Jesus G, Francischetti Ivo M B, Pham Van My, Garfield Mark K, Mather Thomas N, Ribeiro José M C
Medical Entomology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0425, USA.
J Exp Biol. 2002 Sep;205(Pt 18):2843-64. doi: 10.1242/jeb.205.18.2843.
To attempt description of the set of mRNA and protein (sialome) expressed in the salivary glands of the tick Ixodes scapularis, we randomly sequenced 735 clones of a full-length salivary gland cDNA library of this arthropod and performed Edman degradation of protein bands from salivary gland homogenates (SGH) and saliva separated by SDS-PAGE. The sequences were grouped into 410 clusters, of which 383 are not associated with known I. scapularis sequences. 15- and 17-protein bands from PAGE yielded amino-terminal information on the saliva and salivary gland gels, respectively. We attributed 19 of these sequences to translation products of the cDNA library. Full-length sequences were obtained for 87 clones. Among these protein sequences are several protease inhibitors of distinct classes, metalloproteases, novel proteins with histamine-binding domains, and several peptide families of unknown function displaying different conserved cysteine residues, many of which contain single Kunitz domains. This work provides information into the diversity of messages expressed in the salivary glands of I. scapularis, describes novel sequences that may be responsible for known biological activites, indicates further biological activities that may be present in I. scapularis saliva and identifies novel vaccine targets that may be used in Lyme disease prevention.
为了尝试描述肩突硬蜱唾液腺中表达的mRNA和蛋白质(唾液蛋白质组)集合,我们对该节肢动物唾液腺全长cDNA文库的735个克隆进行了随机测序,并对通过SDS-PAGE分离的唾液腺匀浆(SGH)和唾液中的蛋白条带进行了埃德曼降解。这些序列被分为410个簇,其中383个与已知的肩突硬蜱序列无关。PAGE中的15条和17条蛋白条带分别提供了唾液和唾液腺凝胶的氨基末端信息。我们将其中19个序列归因于cDNA文库的翻译产物。获得了87个克隆的全长序列。在这些蛋白质序列中,有几种不同类型的蛋白酶抑制剂、金属蛋白酶、具有组胺结合域的新蛋白质,以及几个功能未知的肽家族,它们显示出不同的保守半胱氨酸残基,其中许多含有单个Kunitz结构域。这项工作提供了有关肩突硬蜱唾液腺中表达信息多样性的信息,描述了可能负责已知生物活性的新序列,指出了肩突硬蜱唾液中可能存在的其他生物活性,并确定了可用于预防莱姆病的新疫苗靶点。