Lactoferrin: similarity to diamine oxidase and purification by aminohexyl affinity chromatography.
A protein reacting with a monoclonal antibody against human placental diamine oxidase was purified from the specific granules of human neutrofil granulocytes using affinity chromatography on aminohexyl-divinylsulfonyl-agarose. The protein had an M(r) determined by SDS/PAGE, corresponding to diamine oxidase, but had other properties which indicated that it might be a different protein. A combination of protein chemical techniques, including N-terminal sequencing, identified the protein as lactoferrin, an iron-containing protein with an M(r) of approximately 800000, a high isoelectric point and ferroxidase activity. Purified commercial lactoferrin was shown to bind to aminohexyl-divinylsulfonyl-agarose, and to be eluted in a heterogenous way from the matrix by amines and salt. Alignment of the sequences of diamine oxidase and lactoferrin showed that they are similar, indicating a common ancestry for these two different classes of metallo-oxidases.
Houen
G
G
Department of Autoimmunology, Statens Serum Institut, Copenhagen, Denmark.
Högdall
E V
EV
Barkholt
V
V
Nørskov
L
L
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry* / Q000276:immunology; D000588:Amines / Q000378:metabolism; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003959:Diamines / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007781:Lactoferrin / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000737:chemistry; D010920:Placenta / Q000201:enzymology; D012685:Sepharose / Q000031:analogs & derivatives / Q000378:metabolism; D016415:Sequence Alignment; D017421:Sequence Analysis
D000588:Amines; D000911:Antibodies, Monoclonal; D003959:Diamines; C008697:octamethylenediamine; C024903:aminohexyl-sepharose; D012685:Sepharose; D006631:Amine Oxidase (Copper-Containing); D007781:Lactoferrin; C008699:octylamine