Characterization of a hyaluronic acid-binding protein from sheep brain comparison with human brain hyaluronectin.
1. A hyaluronic acid (HA)-binding glycoprotein from sheep brain was characterized. 2. The specific affinity for HA was shown in vitro by high performance liquid chromatography, polyacrylamide gel electrophoresis and ELISA methods. 3. The KD for high molecular weight HA was 5.4 10(-9) M at 37 degrees C and lower than 10(-10) M at 4 degrees C. 4. No link protein was found and HA molecules could bind up to 10 times their weight of the glycoprotein. 5. The specific site for interaction was the HA-derived decasaccharide HA10. 6. The protein is composed of one polypeptidic chain. Tryptophan and lysine play a prominent role in the conformation of the binding site to HA. 7. Enzyme analysis indicated that the protein different forms are due to differences in glycosylation and that N- and O-linkages coexist in the molecules. 8. Immunohistochemistry localized the glycoprotein at the nodes of Ranvier and at the periphery of neurons. The perineuronal labeling was seen around all neurons studied in the cerebellum whereas it was almost undetectable in the cerebral hemispheres. 9. HA is not saturated by hyaluronectin (HN) in the sheep nervous system. 10. The glycoprotein is largely similar to human brain HN, and different from the hyaluronate-binding protein characterized in the cartilage.
Delpech
B
B
Laboratoire d'Oncologie moléculaire, Centre Henri-Becquerel, Rouen, France.
Maingonnat
C
C
Delpech
A
A
Maes
P
P
Girard
N
N
Bertrand
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001923:Brain Chemistry; D002236:Carbohydrate Conformation; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000378:metabolism; D015151:Immunoblotting; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012756:Sheep
D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid