This study is the first to report the presence of CD13/glycoprotein 150 (GP150)/aminopeptidase-N activity in cell-free plasma. We have determined that aminopeptidase-N in plasma provides, quantitatively, aminopeptidase-N's predominant functional activity within flowing blood. Thus, while aminopeptidase-N activity observed in whole blood can be partly, but significantly, blocked by the CD13 monoclonal antibody (MAB) WM15, the magnitude of such inhibition is low (< 25%) and similar to that observed using washed cell fractions selectively enriched for neutrophils (30.6% inhibition) or monocytes (21.8% inhibition). Plasma, free of cell components, possesses substantial aminopeptidase-N activity that is largely inhibitable (> 70%) by WM15. Blood collected into heparin or citrate yields similar data, while blood collected into EDTA gives rise to reduced CD13/aminopeptidase-N activity, consistent with inhibition of the known heavy-metal ion association necessary for proper functioning of this molecule. Although monocyte- and granulocyte-enriched cell fractions possess aminopeptidase-N activity significantly inhibitable by CD13 antibodies, lymphocyte-enriched cell fractions also possess aminopeptidase-N-like activity; however, in the latter case, this activity is not inhibitable by CD13 antibodies. Immunoaffinity isolation of plasma aminopeptidase-N has also been carried out; further characterization using functional studies and sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) electrophoresis indicates that CD13 MABs can completely clear plasma of aminopeptidase-N activity and that the purified protein has similar electrophoretic characteristics to cell-derived material. These data, therefore, provide evidence for the presence within blood both of a soluble (that is, non-cell-associated) form of CD13/GP150/aminopeptidase-N localizable to plasma and of cell-associated, aminopeptidase-N-like proteins other than CD13/GP150. These findings have significant implications for our understanding of the many functions of this molecule in blood.
|Number of pages||7|
|Publication status||Published - Dec 1993|