Complex formation between PSA isoenzymes and protease inhibitors.
OBJECTIVE
We have studied complex formation between the isoenzymes of prostate specific antigen (PSA) and protease inhibitors in vitro.


METHODS
Ion exchange chromatography and hydrophobic interaction chromatography (HIC) were used for rapid separation of PSA isoenzymes from inhibitors and for characterization of the complex formation. Immunofluorometric assays (IFMA) specific for free PSA, the PSA-alpha 1-antichymotrypsin (PSA-ACT) complex and for both of these (total PSA) were used to measure various forms of PSA. Loss of free PSA immunoreactivity was used to estimate complex formation with alpha 2-macroglobulin (A2M) and ACT, which also was measured by PSA-ACT IFMA.


RESULTS
Complex formation between PSA and A2M was more rapid than with ACT. After extended incubation, about 75% of PSA reacted with ACT and 85% with A2M. When added to a mixture of ACT and A2M at concentrations corresponding to those in plasma, only 17% of PSA formed a complex with ACT while 17% remained free and 66% was undetectable, indicating complex formation with A2M. After extended incubation of PSA-ACT at 37C, a significant proportion of PSA was released as free active PSA. When A2M was included in the reaction mixture, the loss of PSA-ACT was not accompanied by appearance of free PSA, an indication that it complexed with A2M. Five to 18% of nicked PSA complexed with ACT whereas 54 to 67% reacted with A2M.


CONCLUSIONS
alpha 2-macroglobulin is the major inhibitor of PSA when it reached the circulation. Contrary to earlier assumptions, nicked PSA can bind to A2M rendering it inaccessible to antibodies.
Leinonen
J
J
Department of Clinical Chemistry, Helsinki University Central Hospital, Finland.
Zhang
W M
WM
Stenman
U H
UH
D002845:Chromatography; D015200:Fluoroimmunoassay; D006801:Humans; D007527:Isoenzymes / Q000502:physiology*; D017430:Prostate-Specific Antigen / Q000502:physiology*; D015842:Serine Proteinase Inhibitors / Q000502:physiology*; D000514:alpha 1-Antichymotrypsin / Q000502:physiology*; D000511:alpha-Macroglobulins / Q000502:physiology*
D007527:Isoenzymes; D015842:Serine Proteinase Inhibitors; D000514:alpha 1-Antichymotrypsin; D000511:alpha-Macroglobulins; D017430:Prostate-Specific Antigen