A method for detecting hyaluronan (HA)-binding proteins in transblot assays using biotinylated HA (BHA) is described. Some of the binding characteristics of a novel HA receptor termed RHAMM (Receptor for HA-Mediated Motility) are characterized using this assay. The method is also used to detect other HA-binding proteins in tissue homogenates. This method is semiquantitative, rapid, reproducible, sensitive and therefore of potential use in identifying the levels of HA-binding proteins in different cells and tissues.

A cloned 2184-bp cDNA coding for human interstitial retinol-binding protein (IRBP) has been isolated and sequenced. The probe hybridized to a 5.2-kb poly(A) RNA from human retinas. Nineteen tryptic peptides (363 amino acids) sequenced and purified from bovine IRBP could be aligned with 86-88% homology to the translated sequence. Two segments approximately 200 amino acids long were found to have a 41% residue identity, suggesting an internal duplication event. This cloned cDNA was used to probe DNA samples from a panel of 29 rodent-human somatic cell hybrids, mapping the structural gene for IRBP to chromosome 10. In situ hybridization suggested a regional localization near the centromere (p11.2----q11.2), although a secondary site of hybridization at q24----25 was also observed.

SOURCE/DESCRIPTION: A 2184 bp cDNA (H.4 IRBP) encoding human interstitial retinol-binding protein (IRBP; Fong et al., 1984) isolated from a human retina cDNA library in XgtlO by screening with a bovine IRBP cDNA probe (B.23; Liou et al., 1986).

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