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Expression of Pro Form of Prostate-specific Antigen by Mammalian Cells and Its Conversion to Mature, Active Form by Human Kallikrein 2

Hybritech Incorporated, San Diego, California 92196-9006

To study the expression, biosynthesis, and processing of prostate-specific antigen (PSA) in mammalian cells, recombinant PSA was expressed in Syrian hamster tumor cell line, AV12-664 (AV12-PSA). Expression of PSA was monitored by the Tandem-MP PSA assay. PSA was secreted into the medium during the logarithmic phase of cell growth at > 9 µg/ml and was stable. The PSA purified from spent medium of AV12-PSA cells did not exhibit any enzymatic activity and did not complex with the protease inhibitor, -1-antichymotrypsin. These findings indicated that an inactive form of PSA was expressed by AV12-PSA cells. NH₂-terminal sequencing confirmed the identity of the PSA purified from the spent medium of AV12-PSA cells to be pro-PSA. This demonstrates that PSA is expressed as pro-PSA by mammalian cells and suggests that pro-PSA may be present in biological fluids. Human kallikrein 2 (hK2), another member of the hK family, is also expressed predominantly in prostate epithelium. Although hK2 has been shown to exhibit trypsin-like activity, little is known about its natural substrates. Using purified proteins, we show that hK2 can convert pro-PSA to mature, enzymatically active PSA, thus establishing a physiological connection between hK2 and PSA. These findings imply that hK2 may be regulating PSA activity in vivo.

¹ To whom requests for reprints should be addressed, at Hybritech Incorporated, P. O. Box 269006, San Diego, CA 92196-9006. Phone: (619) 621-3258; Fax: (619) 621-4610; E-mail: mssaedi@beckman.com.

Received 5/ 6/97. Accepted 6/10/97.

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