Structural characterisation of the hepatitis C envelope glycoprotein E1 ectodomain derived from a mammalian and a yeast expression system.
The structure of the ectodomain of the hepatitis C envelope glycoprotein E1 (E1s) was characterised by spectroscopic methods. Monomeric E1s was purified from a mammalian and from a Hansenula polymorpha cell lysate, and cysteine-blocked monomers were reconstituted into stable particles. Particles from yeast E1s and mammalian E1s showed a comparable reactivity in ELISA with sera from human chronic HCV carriers, similar antibody titers in the sera of immunised mice as well as a comparable structure as analyzed by spectroscopic methods (tryptophan fluorescence, circular dichroism, and Fourier transform infrared spectroscopy). The overall secondary structure of E1s was neither influenced by the degree of glycosylation nor by the nature of cysteine modification used during purification. The structural comparability of mammalian- and H. polymorpha-expressed E1s opens new perspectives for further development of E1s-based therapeutics as yeast systems generally allow a more easy scaling up.
Lorent
Eric
E
Laboratory of Biomolecular Dynamics, KULeuven, Celestijnenlaan 200G, 3001 Leuven, Belgium.
Bierau
Horst
H
Engelborghs
Yves
Y
Verheyden
Gert
G
Bosman
Fons
F
D000818:Animals; D002522:Chlorocebus aethiops; D002942:Circular Dichroism; D016174:Hepacivirus / Q000276:immunology; D019698:Hepatitis C, Chronic / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism*; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure
C087035:E1 protein, Hepatitis C virus; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins