Identification of Hyaluronan Binding Proteins Using a Biotinylated Hyaluronan Probe

Hoare, Kishalay; Savani, Rashmin C.; Wang, C.; Yang, Baichun; Turley, Eva A.

doi:10.3109/03008209309041327

Cited by 26 publications

(25 citation statements)

References 23 publications

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“…The blot was incubated with either anti-peptide 168-288 Ab or anti-FP Ab overnight at 4°C and then further probed with biotinylated HA as described previously (29). Streptavidin conjugated with horse radish peroxidase was used to identify bound biotinylated HA.…”

Section: Methodsmentioning

confidence: 99%

Migration of bovine aortic smooth muscle cells after wounding injury. The role of hyaluronan and RHAMM.

Savani¹,

Wang²,

Yang³

et al. 1995

J. Clin. Invest.

191

160

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The migration of smooth muscle cells is a critical event in the pathogenesis of vascular diseases. We have investigated the role of hyaluronan (HA) and the hyaluronan receptor RHAMM in the migration of adult bovine aortic smooth muscle cells (BASMC). Cultured BASMC migrated from the leading edge of a single scratch wound with increased velocity between 1 and 24 h. Polyclonal anti-RHAMM antisera that block HA binding with this receptor abolished smooth muscle cell migration following injury. HA stimulated the random locomotion of BASMC and its association with the cell monolayer increased following wounding injury. Immunoblot analysis of wounded monolayers demonstrated a novel RHAMM protein isoform that appeared within one hour after injury. At the time of increased cell motility after wounding, FACS® analysis demonstrated an increase in the membrane localization in -25% of the cell population. Confocal microscopy of injured monolayers confirmed that membrane expression of this receptor was limited to cells at the wound edge. Collectively, these data demonstrate that RHAMM is necessary for the migration of smooth muscle cells and that expression and distribution of this receptor is tightly regulated following wounding of BASMC monolayers. (J. Clin. Invest. 1995Invest. . 95:1158Invest. -1168

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Section: Methodsmentioning

confidence: 99%

Migration of bovine aortic smooth muscle cells after wounding injury. The role of hyaluronan and RHAMM.

Savani¹,

Wang²,

Yang³

et al. 1995

J. Clin. Invest.

191

160

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“…Analysis of primary astrocytes at various times after sparse plating indicated a progressive decrease in RHAMM expression as cultures became confluent (data not shown). In immunoblots stripped of antibody and reprobed with biotinylated-HA for detection of proteins having HA binding capacity (Hoare et al, 1993), primary astrocytes were found to produce three major HA binding proteins one of which corresponded to the 72 kDa protein recognized by anti-RHAMM antibody. The identity of the other two HA binding proteins that had molecular weights of 140 and 50 kDa and did not react with RHAMM antibody are unknown.…”

Section: Expression Of Rhamm In Glial Cellsmentioning

confidence: 99%

“…HA binding of electrophoresed proteins was analyzed according to previously published protocols using biotinylated HA as a probe. The specificity of this technique has previously been established by showing that digestion of biotinylated HA with streptomyces hyaluronidase abolishes binding and that binding is inhibited by excess unlabelled HA (Hoare et al, 1993). For this assay, immunoblots that had been probed with anti-RHAMM antibody were stripped as previously described (Hall et al, 1993(Hall et al, , 1994 and membranes were then incubated with biotinylated HA (0.001 pg/ml) for 1 h, washed, and developed for chemiluminescence using luminol as described (Amersham, UK).…”

Section: Western Blot and Hyaluronan Binding Analysesmentioning

confidence: 99%

Astrocyte and microglial motility in vitro is functionally dependent on the hyaluronan receptor RHAMM

Turley

Hossain

Sorokan

et al. 1994

Glia

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RHAMM (Receptor for Hyaluronic Acid Mediated Motility) has been identified as a receptor for the extracellular matrix component hyaluronan (HA) and was recently shown to be essential for the locomotion of normal and transformed peripheral cells. Until now the potential role of RHAMM in the motility of neural-derived cells has not been investigated. Here, we report that cultured primary astrocytes, astrocyte cell lines, and microglia express this receptor and exhibit RHAMM-dependent motility. Immunocytochemical localization of RHAMM showed that it was often present as aggregates at the periphery of cells in contact with one another or concentrated on protruding processes of isolated cells. Glial cells contained 50 and 72 kDa forms of RHAMM, and both of these forms were found to have HA binding capacity. Time lapse imaging of cell locomotion revealed a significant inhibition of motility and process elongation by neutralizing anti-RHAMM antibodies and by peptides corresponding to the HA binding domains of RHAMM. These results demonstrate that RHAMM serves a role in glial cell locomotion in vitro and provide the basis for investigations of the motile behavior of glial cells in vivo after CNS injury.

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“…Furthermore, most malignant solid tumors and their surrounding stromal tissue contain elevated levels of HA (9). CD44 is the principal cell surface receptor for HA, and it presents at low levels on epithelial, hemopoietic and some neuronal cells (11)(12)(13), and at elevated levels in various carcinoma, lymphoma, breast, colorectal, and lung tumor cells (14)(15)(16)(17). Overexpression of the HA receptors CD44 and RHAMM on cancer cells results in enhanced binding and endocytosis uptake of such compound.…”

Section: Introductionmentioning

confidence: 99%

Intratumoral Delivery of Paclitaxel in Solid Tumor from Biodegradable Hyaluronan Nanoparticle Formulations

et al. 2009

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Abstract. In the current study, novel paclitaxel-loaded cross-linked hyaluronan nanoparticles were engineered for the local delivery of paclitaxel as a prototype drug for cancer therapy. The nanoparticles were prepared using a desolvation method with polymer cross-linking. In vitro cytotoxicity studies demonstrated that less than 75% of the MDA-MB-231 and ZR-75-1 breast cancer cells were viable after 2-day exposure to paclitaxel-loaded hyaluronan nanoparticles or free paclitaxel, regardless of the dose. These results suggest that hyaluronan nanoparticles maintain the pharmacological activity of paclitaxel and efficiently deliver it to the cells. Furthermore, in vivo administration of the drug-loaded nanoparticles via direct intratumoral injection to 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumor in female rats was studied. The paclitaxel-loaded nanoparticles treated group showed effective inhibition of tumor growth in all treated rats. Interestingly, there was one case of complete remission of tumor nodule and two cases of persistent reduction of tumor size that was observed on subsequent days. In the case of free paclitaxel-treated group, the mean tumor volume increased almost linearly (R 2 =0.93) with time to a size that was 4.9-fold larger than the baseline volume at 57 days post-drug administration. Intratumoral administration of paclitaxel-loaded hyaluronan nanoparticles could be a promising treatment modality for solid mammary tumors.

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Identification of Hyaluronan Binding Proteins Using a Biotinylated Hyaluronan Probe

Cited by 26 publications

References 23 publications

Migration of bovine aortic smooth muscle cells after wounding injury. The role of hyaluronan and RHAMM.

Migration of bovine aortic smooth muscle cells after wounding injury. The role of hyaluronan and RHAMM.

Astrocyte and microglial motility in vitro is functionally dependent on the hyaluronan receptor RHAMM

Intratumoral Delivery of Paclitaxel in Solid Tumor from Biodegradable Hyaluronan Nanoparticle Formulations

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