The detection of bovine lactoferrin binding protein on Trypanosoma brucei.
Trypanosoma brucei, the causative agent of sleeping sickness in humans, requires transferrin (TF) for growth. Therefore, T. brucei has a TF receptor that allows it to obtain iron from TF. Lactoferrin (LF), a member of the TF family protein, is an iron-binding protein that is found in most biological fluids of mammals. LF has been shown to interact with some bacteria species by specific receptor-ligand binding. We examined the ability of T. brucei to bind bovine LF (bLF) by using a fluorescence test, streptavidin-biotin (SAB) microplate analysis, and far Western blotting using a biotin-streptavidin system. We found that bLF bound to components of T. brucei, and that bLF hydrolysate disrupted the sites responsible for binding to parasite proteins. Furthermore, bLF, human LF, bovine TF, and ovotransferrin bound same proteins of T. brucei, which exhibited molecular masses of 40 and 43 kDa. The N-terminal amino acid sequence of the 40 kDa bLF binding protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH).
Tanaka
Tetsuya
T
Dairy Science Laboratory, Graduate School of Agriculture, Hokkaido University, Japan.
Abe
Yukie
Y
Inoue
Noboru
N
Kim
Woan-Sub
WS
Kumura
Haruto
H
Nagasawa
Hideyuki
H
Igarashi
Ikuo
I
Shimazaki
Kei-Ichi
K
D000818:Animals; D026481:Blotting, Far-Western / Q000662:veterinary; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D003207:Conalbumin / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000378:metabolism*
D002352:Carrier Proteins; C490098:LTF protein, human; D015800:Protozoan Proteins; D014168:Transferrin; C086010:glycyrrhizin-binding proteins; D003207:Conalbumin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007781:Lactoferrin