1887

Abstract

The human immunodeficiency virus (HIV) type 1 gene product was expressed as an N-terminal fusion protein with glutathione--transferase (GST) in the baculovirus system. The resulting GST fusion protein was found to be authentically myristylated at the N terminus and could be purified to homogeneity by one-step affinity chromatography on immobilized glutathione. The high affinity of GST for glutathione was exploited to develop an assay to identify cellular proteins capable of interacting with . Several such proteins were identified in extracts from the Jurkat human T cell line. The interaction between -binding proteins and immobilized GST could be specifically competed by the addition of soluble . Cell fractionation showed that -binding proteins were present in both cytosolic and membrane-associated fractions. A non-myristylated derivative failed to bind to the membrane-associated proteins but was able to bind to the cytosolic group, albeit with reduced affinity. In addition, a single protein present in both soluble and membrane-associated fractions exhibited myristylation-independent binding to . By analogy with other myristylated proteins such as MARCKS (myristylated alanine-rich C kinase substrate) and the Rous sarcoma virus transforming protein, , the membrane-associated proteins that bind only to myristylated may represent a specific membrane target for . The cytosolic proteins that interact with may constitute soluble components of an as yet unidentified signal transduction pathway which is the target of action in the HIV-1-infected cell.

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1993-08-01
2021-10-17
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