Complement factor H is a serum-binding protein for adrenomedullin, and the resulting complex modulates the bioactivities of both partners.
Adrenomedullin (AM) is an important regulatory peptide involved in both physiological and pathological states. We have previously demonstrated the existence of a specific AM-binding protein (AMBP-1) in human plasma. In the present study, we developed a nonradioactive ligand blotting assay, which, together with high pressure liquid chromatography/SDS-polyacrylamide gel electrophoresis purification techniques, allowed us to isolate AMBP-1 to homogeneity. The purified protein was identified as human complement factor H. We show that AM/factor H interaction interferes with the established methodology for quantification of circulating AM. Our data suggest that this routine procedure does not take into account the AM bound to its binding protein. In addition, we show that factor H affects AM in vitro functions. It enhances AM-mediated induction of cAMP in fibroblasts, augments the AM-mediated growth of a cancer cell line, and suppresses the bactericidal capability of AM on Escherichia coli. Reciprocally, AM influences the complement regulatory function of factor H by enhancing the cleavage of C3b via factor I. In summary, we report on a potentially new regulatory mechanism of AM biology, the influence of factor H on radioimmunoassay quantification of AM, and the possible involvement of AM as a regulator of the complement cascade.
Pio
R
R
Department of Cell and Cancer Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Martinez
A
A
Unsworth
E J
EJ
Kowalak
J A
JA
Bengoechea
J A
JA
Zipfel
P F
PF
Elsasser
T H
TH
Cuttitta
F
F
D053607:Adrenomedullin; D000818:Animals; D001769:Blood; D015153:Blotting, Western; D002455:Cell Division / Q000502:physiology; D002851:Chromatography, High Pressure Liquid; D017242:Complement Factor H / Q000378:metabolism* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010455:Peptides / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D014407:Tumor Cells, Cultured
D053607:Adrenomedullin; D017242:Complement Factor H; D010455:Peptides; C493495:CFH protein, human