Identification of dengue virus binding proteins using affinity chromatography.

Area of Science:

• Virology
• Biochemistry
• Molecular Biology

Background:

• Virus overlay protein binding assays (VOPBA) have identified potential dengue virus receptors.
• VOPBA involves protein denaturation during electrophoresis, raising questions about the physiological relevance of identified proteins.

Purpose of the Study:

• To develop a more physiologically relevant method for identifying dengue virus binding proteins.
• To validate findings from VOPBA studies by using a technique that minimizes protein denaturation.

Main Methods:

• Construction of a Sepharose 4B-dengue virus serotype 2-affinity column.
• Selective binding of dengue virus binding proteins from HepG2 (liver) cell membrane preparations.

Main Results:

• Glucose-Regulated Protein 78 (GRP78) was specifically bound by the affinity column.
• The 37/67 kDa high affinity laminin receptor was not specifically bound.
• Findings suggest GRP78 is a potential dengue virus binding protein.

Conclusions:

• Affinity chromatography is a valuable adjunct to VOPBA for identifying physiologically relevant viral receptors.
• Proteins identified through VOPBA can be physiologically relevant, but affinity chromatography offers a more reliable approach for denaturation-sensitive proteins.