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First published online September 15, 2004
Journal of Experimental Biology 207, 3717-3729 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.01185

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Identification of the most abundant secreted proteins from the salivary glands of the sand fly Lutzomyia longipalpis, vector of Leishmania chagasi

Jesus G. Valenzuela^1,*, Mark Garfield², Edgar D. Rowton³ and Van M. Pham¹

¹ Vector Molecular Biology Unit, Laboratory of Malaria and Vector Research, NIAID, National Institutes of Health, 12735 Twinbrook Parkway, Room 2E-22C, Rockville, MD 20852, USA,
² Structural Biology Unit, NIAID, Rockville, MD 20852, USA
³ Department of Entomology, Walter Reed Army Institute of Research, Washington, DC 20307, USA

^* Author for correspondence (e-mail: jvalenzuela{at}niaid.nih.gov)

Accepted 12 July 2004

Using massive cDNA sequencing, proteomics and customized computational biology approaches, we have isolated and identified the most abundant secreted proteins from the salivary glands of the sand fly Lutzomyia longipalpis. Out of 550 randomly isolated clones from a full-length salivary gland cDNA library, we found 143 clusters or families of related proteins. Out of these 143 families, 35 were predicted to be secreted proteins. We confirmed, by Edman degradation of Lu. longipalpis salivary proteins, the presence of 17 proteins from this group. Full-length sequence for 35 cDNA messages for secretory proteins is reported, including an RGD-containing peptide, three members of the yellow-related family of proteins, maxadilan, a PpSP15-related protein, six members of a family of putative anticoagulants, an antigen 5-related protein, a D7-related protein, a cDNA belonging to the Cimex apyrase family of proteins, a protein homologous to a silk protein with amino acid repeats resembling extracellular matrix proteins, a 5'-nucleotidase, a peptidase, a palmitoyl-hydrolase, an endonuclease, nine novel peptides and four different groups of proteins with no homologies to any protein deposited in accessible databases. Sixteen of these proteins appear to be unique to sand flies. With this approach, we have tripled the number of isolated secretory proteins from this sand fly. Because of the relationship between the vertebrate host immune response to salivary proteins and protection to parasite infection, these proteins are promising markers for vector exposure and attractive targets for vaccine development to control Leishmania chagasi infection.

Key words: salivary gland transcript, salivary protein, New World sand fly, Lutzomyia longipalpis, saliva

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