Isolation of glycoprotein D from herpes simplex virus type 1 by gel filtration high performance liquid chromatography.
Rabbit kidney (RK-13) and human jejunum and ileum (I-407) cells infected with herpes simplex virus type 1, strain F, were radiolabelled with [14C]glucosamine or [35S]methionine for 24 h. The cells were extracted with 1% Triton X-100 and the extracts were separated by gel filtration high performance liquid chromatography. Monoclonal antibody immunoprecipitation of the fractions collected from the column revealed a monomeric glycoprotein D (gD) of 52 - 56,000 molecular weight from RK-13 cells and two monomeric forms of gD, 54,000 and 58,000 molecular weight, from I-407 cells. Densitometry scanning of the autoradiograms from SDS-PAGE showed gD from the RK-13 host cells to be 98.7% pure with the [35S]methionine label and 97.0% pure with the [14C]glucosamine. On the other hand, gD from the I-407 host cells was only 78.6% with the [35S]methionine label and 96% pure with the [14C]glucosamine. This method could provide a means for the isolation of native gD for structural and immunological studies.
McGarry
T J
TJ
Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia.
al-Ahdal
M N
MN
D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007082:Ileum / Q000032:analysis; D007583:Jejunum / Q000032:analysis; D007668:Kidney / Q000032:analysis; D011817:Rabbits; D018139:Simplexvirus; D014759:Viral Envelope Proteins / Q000302:isolation & purification*
D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1