Bh. Yang et al., BIOTINYLATED HYALURONIC-ACID AS A PROBE FOR IDENTIFYING HYALURONIC ACID-BINDING PROTEINS, Analytical biochemistry, 228(2), 1995, pp. 299-306
The glycosaminoglycans hyaluronan (HA), heparin, and chondroitin sulfa
te were biotinylated using biotin-x-hhydrazide (biotin-epsilon-aminoca
proyl hydrozyde) in conjunction with N-ethyl-N'-(3-dimethylaminopropyl
)carbodiimide hydrochloride, an activating agent for carboxyl groups.
The biotin-x-hydrazide was shown to be coupled directly to the glycosa
minoglycans in enzymatic digestions and competition experiments. The b
iotinylated HA was shown to bind to link protein and receptor for hyal
uronic acid-mediated motility, two proteins known to bind HA. The labe
led HA was used as a probe to detect known HA-binding proteins in chic
ken cartilage extract and to identify new HA-binding motifs in the G3
domain of the proteoglycan aggrecan. The significance of the biotinyla
tion of HA, heparin, and chondroitin sulfate A are discussed. (C) 1995
Academic Press, Inc.