A putative serine protease among the excretory-secretory glycoproteins of L1 Trichinella spiralis.
Trichinella spiralis first-stage larvae infect susceptible hosts by invading epithelial cells that line the small intestine. During this process the larva disgorges several glycoproteins that bear an unusual, highly antigenic sugar moiety, tyvelose (3,6-dideoxy arabinohexose). Monoclonal antibodies specific for tyvelose protect the intestine against infection, implicating tyvelose-bearing glycoproteins as mediators of invasion and niche establishment in the intestinal epithelium. In order to investigate these glycoproteins at the molecular level, we first prepared monoclonal anti-peptide antibodies. The antibodies bind a family of glycoproteins that are present in excretory-secretory products of first-stage larvae and are delivered to epithelial cells during invasion by T. spiralis. The major species present in an affinity purified fraction of crude T. spiralis antigens were subjected to tryptic peptide digestion. De novo amino acid sequencing of the peptides using Q-TOF tandem mass spectrometry, in combination with database searches and antibody screening of an L1 cDNA library, showed that the glycoproteins are variably glycosylated homologues of the serine protease family.
Romaris
Fernanda
F
James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
North
Simon J
SJ
Gagliardo
Lucille F
LF
Butcher
Barbara A
BA
Ghosh
Kaya
K
Beiting
Daniel P
DP
Panico
Maria
M
Arasu
Prema
P
Dell
Anne
A
Morris
Howard R
HR
Appleton
Judith A
JA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D004847:Epithelial Cells / Q000469:parasitology; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006601:Hexoses / Q000276:immunology; D007814:Larva / Q000201:enzymology / Q000254:growth & development / Q000472:pathogenicity; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D051381:Rats; D020539:Sequence Analysis, Protein; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017160:Trichinella spiralis / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity
D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D018076:DNA, Complementary; D006023:Glycoproteins; D006601:Hexoses; D010455:Peptides; C009616:tyvelose; D012697:Serine Endopeptidases