The Tat protein has several functional domains, one of which is the cysteine-rich domain that is a highly conserved region in spite of the presence of many subtypes of human immunodeficiency virus type 1 (HIV-1). Although the cysteine-rich domain is a potential site for Zn(2+) binding, it is controversial whether Zn(2+) is substantially essential for the structure and activities of the Tat protein. To study the significance of Zn(2+) in the cysteine-rich domain of the Tat protein particularly released to the extracellular space, we raised the monoclonal antibody (MAb) 5A4, which has an attractive property of recognizing the Zn(2+)-binding Tat(20-41) peptide but not the apo-Tat(20-41) peptide.
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