Addition of purified prostate specific antigen to serum from female subjects: studies on the relative inhibition by alpha 2-macroglobulin and alpha 1-antichymotrypsin.
OBJECTIVE
Two forms of prostate specific antigen (PSA), 1 complexed to alpha 1-antichymotrypsin and the other a free PSA, are recognized by current commercially available immunoassays. A third form of PSA complexed to alpha 2-macroglobulin also is present in the serum. To study these 3 different molecular forms of PSA in vivo, we simulated leakage of PSA from the prostate into the serum in vitro.


METHODS
Purified seminal fluid PSA was incubated with fresh sera from female subjects at different concentrations. Following gel filtration chromatography, the 3 forms of PSA were studied by immunoassays and Western blot analysis.


RESULTS
Using a commercial immunoassay, 60% of immunoreactivity of seminal fluid PSA was lost after incubation with sera from female subjects. Western blot analysis showed that most of this loss in PSA signal was caused by complexation to alpha 2-macroglobulin. Minimal, if any, complexation to alpha 1-antichymotrypsin occurred even when excess alpha 1-antichymotrypsin was added to the serum.


CONCLUSIONS
Our studies demonstrated that alpha 2-macroglobulin is a much stronger inhibitor to PSA than alpha 1-antichymotrypsin. Further studies of these complexes may be important. They clearly explain why spiking PSA into sera from female subjects to be used as quality controls for PSA assays leads only to the free form of enzymatically inactive PSA in the serum, and not to the dominant form of complexed PSA and alpha 1-antichymotrypsin present in human serum.
Chen
Z
Z
Department of Urology, Stanford University School of Medicine, California, USA.
Komatsu
K
K
Prestigiacomo
A
A
Stamey
T A
TA
D005260:Female; D006801:Humans; D007118:Immunoassay; D017430:Prostate-Specific Antigen / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D000514:alpha 1-Antichymotrypsin / Q000378:metabolism* / Q000494:pharmacology; D000511:alpha-Macroglobulins / Q000378:metabolism* / Q000494:pharmacology
D015842:Serine Proteinase Inhibitors; D000514:alpha 1-Antichymotrypsin; D000511:alpha-Macroglobulins; D017430:Prostate-Specific Antigen