Abstract
The adenovirus fiber protein is used for attachment of the virus to a specific receptor on the cell surface. Structurally, the protein consists of a long, thin shaft that protrudes from the vertex of the virus capsid and terminates in a globular domain termed the knob. To verify that the knob is the domain which interacts with the cellular receptor, we have cloned and expressed the knob from adenovirus type 5 together with a single repeat of the shaft in Escherichia coli. The protein was purified by conventional chromatography and functionally characterized for its interaction with the adenovirus receptor. The recombinant knob domain bound about 4,700 sites per HeLa cell with an affinity of 3 x 109 M-1 and blocked adenovirus infection of human cells. Antibodies raised against the knob also blocked virus infection. By gel filtration and X-ray diffraction analysis of protein crystals, the knob was shown to consist of a homotrimer of 21-kDa subunits. The results confirm that the trimeric knob is the ligand for attachment to the adenovirus receptor.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 5239-5246 |
| Number of pages | 8 |
| Journal | Journal of Virology |
| Volume | 68 |
| Issue number | 8 |
| State | Published - Aug 1994 |
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ASJC Scopus subject areas
- Immunology
Cite this
Characterization of the knob domain of the adenovirus type 5 fiber protein expressed in Escherichia coli. / Henry, Lynda J.; Xia, Di; Wilke, Marjorie E.; Deisenhofer, Johann; Gerard, Robert D.
In: Journal of Virology, Vol. 68, No. 8, 08.1994, p. 5239-5246.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of the knob domain of the adenovirus type 5 fiber protein expressed in Escherichia coli
AU - Henry, Lynda J.
AU - Xia, Di
AU - Wilke, Marjorie E.
AU - Deisenhofer, Johann
AU - Gerard, Robert D.
PY - 1994/8
Y1 - 1994/8
N2 - The adenovirus fiber protein is used for attachment of the virus to a specific receptor on the cell surface. Structurally, the protein consists of a long, thin shaft that protrudes from the vertex of the virus capsid and terminates in a globular domain termed the knob. To verify that the knob is the domain which interacts with the cellular receptor, we have cloned and expressed the knob from adenovirus type 5 together with a single repeat of the shaft in Escherichia coli. The protein was purified by conventional chromatography and functionally characterized for its interaction with the adenovirus receptor. The recombinant knob domain bound about 4,700 sites per HeLa cell with an affinity of 3 x 109 M-1 and blocked adenovirus infection of human cells. Antibodies raised against the knob also blocked virus infection. By gel filtration and X-ray diffraction analysis of protein crystals, the knob was shown to consist of a homotrimer of 21-kDa subunits. The results confirm that the trimeric knob is the ligand for attachment to the adenovirus receptor.
AB - The adenovirus fiber protein is used for attachment of the virus to a specific receptor on the cell surface. Structurally, the protein consists of a long, thin shaft that protrudes from the vertex of the virus capsid and terminates in a globular domain termed the knob. To verify that the knob is the domain which interacts with the cellular receptor, we have cloned and expressed the knob from adenovirus type 5 together with a single repeat of the shaft in Escherichia coli. The protein was purified by conventional chromatography and functionally characterized for its interaction with the adenovirus receptor. The recombinant knob domain bound about 4,700 sites per HeLa cell with an affinity of 3 x 109 M-1 and blocked adenovirus infection of human cells. Antibodies raised against the knob also blocked virus infection. By gel filtration and X-ray diffraction analysis of protein crystals, the knob was shown to consist of a homotrimer of 21-kDa subunits. The results confirm that the trimeric knob is the ligand for attachment to the adenovirus receptor.
UR - http://www.scopus.com/inward/record.url?scp=0028283086&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028283086&partnerID=8YFLogxK
M3 - Article
C2 - 8035520
AN - SCOPUS:0028283086
VL - 68
SP - 5239
EP - 5246
JO - Journal of Virology
JF - Journal of Virology
SN - 0022-538X
IS - 8
ER -