A cDNA clone, B4-2, was isolated from a natural killer (NK) minus T cell subtractive library. The B4-2 clone coded for an mRNA of 2061 bp in length. It encodes a deduced 327 aa protein with a calculated molecular mass of 35.2 kDa. Searching of B4-2 DNA and protein sequences against various databases revealed no high homology to other sequences. However, B4-2 has an unusually high proline content (13%), contains a putative nuclear targeting sequence, and has several SPXX motifs which are frequently found in gene regulatory proteins. One of the stretches of prolines in B4-2 closely resembles the ligand for proteins with SH3 domains. Northern hybridization data showed that B4-2 is not a lymphoid specific gene and is expressed in a hepatoma cell line and also weakly transcribed or absent in a variety of other cells. A polyclonal antiserum raised against recombinant B4-2 recognizes a 32-34 kDa protein in lymphocytes.