ESAG 6 and 7 products of Trypanosoma brucei form a transferrin binding protein complex.
In Trypanosoma brucei, the gene for the expressed variant surface glycoprotein (VSG) is preceded by a series of open reading frames designated expression site associated genes (ESAGs), which together with the VSG gene form a polycistronic transcription unit. It is shown that the products derived from two ESAGs (ESAG 6 and 7 in the nomenclature of Pays, E., et al. Cell 57, 835-845 (1989)) form a complex, which binds transferrin with high affinity. Transferrin affinity chromatography yields heterodimers or higher order heteroligomers composed of the products of ESAG 6 and ESAG 7. The former is a heterogeneously glycosylated protein of 50 to 60 kDa modified by a glycosylphosphatidylinositol membrane anchor at the COOH-terminus, while the latter is the previously identified 42 kDa glycoprotein carrying an unmodified COOH-terminus (Schell, D., et al. EMBO J. 10, 1061-1066 (1991) and Schell, D., et al. EMBO J. 12, 2990 (1993)). When isolated from trypanosomes grown in rodents, the complex is in part free and in part associated with transferrin. Also, the complex is present both in the membrane fraction and the soluble fraction of cell lysates. As shown by immunoelectron microscopy, both transferrin and ESAG 6/7-derived proteins can be demonstrated in the lumen of the flagellar pocket, an invagination of the plasma membrane serving as the sole site for endocytotic uptake of macromolecular nutrients. Weak labeling is also obtained on the flagellar pocket membrane and in intracellular vesicles. The possibility that the binding protein complex serves as a receptor for the uptake of transferrin in T. brucei is discussed.
Steverding
D
D
Max-Planck-Institut für Biologie, Abteilung Membranbiochemie, Tübingen, Germany.
Stierhof
Y D
YD
Chaudhri
M
M
Ligtenberg
M
M
Schell
D
D
Beck-Sickinger
A G
AG
Overath
P
P
D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D006023:Glycoproteins / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000032:analysis; D046911:Macromolecular Substances; D051379:Mice; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000378:metabolism*
C090143:ESAG 6 protein, Trypanosoma brucei; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D046911:Macromolecular Substances; D015800:Protozoan Proteins; D014168:Transferrin; C078011:ESAG 7 protein, Trypanosoma brucei