Purification of membrane-bound lactoferrin from the human milk fat globule membrane.
Although lactoferrin is known as a basic soluble glycoprotein, the presence of the membrane-bound form of this protein has also been demonstrated in human milk. Membrane-bound lactoferrin was extracted from the human milk fat globule membrane with a detergent mixture of 1% Tween-20, 0.5% C12E8, and 0.5 M KCl in 20 mM Tris-HCl (pH 7.4). Lactoferrin in the detergent-soluble fraction was purified by affinity chromatography with Concanavalin A and by hydrophobic chromatography with phenyl-Superose. The purified protein gave a single band of 80 kDa by SDS-PAGE. Its N-terminal amino acid sequence was consistent with that of human lactoferrin.
Cho
J K
JK
Department of Applied Biochemistry, Utsunomiya University, Japan.
Azuma
N
N
Lee
C H
CH
Yu
J H
JH
Kanno
C
C
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003208:Concanavalin A; D005260:Female; D006017:Glycolipids / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D007781:Lactoferrin / Q000032:analysis*; D066292:Lipid Droplets; D008565:Membrane Proteins / Q000032:analysis*; D008895:Milk, Human / Q000737:chemistry*; D008969:Molecular Sequence Data; D012685:Sepharose / Q000031:analogs & derivatives
D006017:Glycolipids; D006023:Glycoproteins; D008565:Membrane Proteins; C110367:milk fat globule; C091883:phenyl-superose; D003208:Concanavalin A; D012685:Sepharose; D007781:Lactoferrin