Identification and molecular characterization of a novel protein Saglin as a target of monoclonal antibodies affecting salivary gland infectivity of Plasmodium sporozoites.
Molecular mechanisms underlying the interaction between malarial sporozoites and putative receptor(s) on the salivary glands of Anopheles gambiae remain largely unknown. In previous studies, a salivary gland protein of ~100 kDa was identified as a putative target based on recognition of the protein by a monoclonal antibody (mAb) 2A3 that caused a >/= 70% reduction in the average number of sporozoites per infected salivary gland when fed to mosquitoes. Using affinity purification we purified the target of this mAb from extracts of female A. gambiae salivary glands and it was found to be a novel protein by tandem mass spectrometric analysis. Biochemical and molecular characterization of the 100 kDa protein showed that this molecule, designated Saglin, exists as a disulphide-bonded homodimer of 50 kDa subunits. The ability to form homodimers was retained even in the recombinant Saglin expressed in mammalian cells (HEK293). The amino acid sequence of Saglin contains a signal peptide suggesting that Saglin is a secreted protein. If Saglin is indeed involved in the process of invasion of A. gambiae salivary glands by sporozoites of Plasmodium, it could provide a novel target for future investigations aimed at interruption of malaria transmission.
Okulate
M A
MA
Department of Molecular Microbiology and Immunology, Johns Hopkins Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.
Kalume
D E
DE
Reddy
R
R
Kristiansen
T
T
Bhattacharyya
M
M
Chaerkady
R
R
Pandey
A
A
Kumar
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000235:genetics* / Q000276:immunology / Q000469:parasitology*; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D017344:Genes, Insect; D006790:Host-Parasite Interactions; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007303:Insect Vectors / Q000235:genetics / Q000276:immunology / Q000469:parasitology; D008288:Malaria / Q000635:transmission; D051379:Mice; D008969:Molecular Sequence Data; D010961:Plasmodium / Q000472:pathogenicity*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012469:Salivary Glands / Q000276:immunology / Q000469:parasitology; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D034101:Sporozoites; D053719:Tandem Mass Spectrometry
D000911:Antibodies, Monoclonal; D019476:Insect Proteins; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D004247:DNA