Visualization of heparin-binding proteins by ligand blotting with /sup 125/I-heparin
A ligand-blotting procedure which allows detection of heparin-binding proteins is described. Crude commercial heparin was fractionated by chromatography on a column of human plasma low-density lipoproteins immobilized to Sepharose CL-4B. Chromatography yielded an unbound and a bound fraction of heparin, designated URH and HRH, respectively. The HRH fraction was reacted with the N-hydroxysuccinimidyl ester of 3-(p-hydroxyphenyl)propionic acid and then labeled with /sup 125/I. Proteins were separated by 3-20% pore-gradient gel electrophoresis, transferred to nitrocellulose, and then assayed for their ability to bind /sup 125/I-labeled HRH. Human plasma apolipoproteins B-100, B-48, and E of chylomicrons, very low-density lipoproteins, and low-density lipoproteins bound the /sup 125/I-labeled HRH; the radiolabeled haparin did not bind to serum albumin, ferritin, catalase, and lactate dehydrogenase. The ligand-blotting procedure should facilitate the purification of heparin-binding domains from these proteins and, moreover may be applicable to the investigation of heparin-protein interactions in general. 15 references.
- Research Organization:
- Univ. of Cincinnati College of Medicine, OH
- OSTI ID:
- 5864005
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 137:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANTICOAGULANTS
APOLIPOPROTEINS
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CELLULOSE ESTERS
CHEMICAL EXPLOSIVES
CHROMATOGRAPHY
CHYLOMICRONS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ESTERS
EXPLOSIVES
HEMATOLOGIC AGENTS
HEPARIN
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
LABELLING
LIGANDS
LIPIDS
LIPOPROTEINS
MUCOPOLYSACCHARIDES
NITRIC ACID ESTERS
NITROCELLULOSE
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
POLYSACCHARIDES
PROTEINS
RADIOASSAY
RADIOISOTOPES
SACCHARIDES
SEPARATION PROCESSES