title: Molecular cloning and characterization of a C-type lectin from Ancylostoma ceylanicum: evidence for a role in hookworm reproductive physiology.
abstract: Lectins comprise a family of related proteins that mediate essential cell functions through binding to carbohydrates. Within this protein family, C-type lectins are defined by the requirement of calcium for optimal biologic activity. Using reverse transcription PCR, a cDNA corresponding to a putative C-type lectin has been amplified from the hookworm parasite Ancylostoma ceylanicum. The 550 nucleotide open reading frame of the A. ceylanicum C-type Lectin-1 (AceCTL-1) cDNA corresponds to a 167 amino acid mature protein (18,706 Da) preceded by a 17 amino acid secretory signal sequence. The recombinant protein (rAceCTL-1) was expressed in Drosophila S2 cells and purified using a combination of affinity chromatography and reverse phase HPLC. Using in vitro carbohydrate binding studies, it was determined that rAceCTL-1 binds N-acetyl-d-glucosamine, a common component of eukaryotic egg cell membranes. Using a polyclonal IgG raised against the recombinant protein, the native AceCTL-1 was identified in sperm and soluble protein extracts of adult male A. ceylanicum by immunoblot. Probing of adult hookworm sections with the polyclonal IgG demonstrated localization to the testes in males, as well as the spermatheca and developing embryos in females, consistent with its role as a sperm protein. Together, these data strongly suggest that AceCTL-1 is a male gender-specific C-type lectin with a function in hookworm reproductive physiology.
authors.affiliation: Program in International Child Health, Department of Pediatrics, Yale University School of Medicine, 464 Congress Avenue, New Haven, CT 06520, USA.
authors.forename: Allison C
authors.initials: AC
authors.lastname: Brown
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Harrison
authors.forename: Wadim
authors.initials: W
authors.lastname: Kapulkin
authors.forename: Brian F
authors.initials: BF
authors.lastname: Jones
authors.forename: Anindita
authors.initials: A
authors.lastname: Sinha
authors.forename: Amy
authors.initials: A
authors.lastname: Savage
authors.forename: Nicholas
authors.initials: N
authors.lastname: Villalon
authors.forename: Michael
authors.initials: M
authors.lastname: Cappello
mesh_terms: D000595:Amino Acid Sequence; D000722:Ancylostoma / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D000724:Ancylostomiasis / Q000469:parasitology*; D000818:Animals; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary; D005260:Female; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D037181:Lectins, C-Type / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008297:Male; D008647:Mesocricetus; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D012098:Reproduction; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D015801:Helminth Proteins; D037181:Lectins, C-Type