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J. Biol. Chem., Vol. 275, Issue 38, 29829-29839, September 22, 2000
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From the Department of Anatomy and Cellular Biology, Tufts
University School of Medicine, Boston, Massachusetts 02111
Hyaluronan has well defined functions in
extracellular matrices and at the surface of cells. However, several
studies have now shown that significant pools of hyaluronan are also
present intracellularly, but its function therein is unknown. One
avenue of investigation that may assist in defining the function of
intracellular hyaluronan is to identify intracellular
hyaluronan-binding proteins. In previous studies we identified CDC37, a
cell cycle regulatory protein, using a monoclonal antibody that
recognizes a novel group of hyaluronan-binding proteins. In this study,
we have identified a second hyaluronan-binding protein with this
antibody and characterized its properties. This protein, which we have
termed IHABP4, was also found to be an intracellular and a specific
hyaluronan-binding protein, containing several hyaluronan-binding
motifs: (R/K)X7(R/K) (where R/K denotes
arginine or lysine and X denotes non-acidic amino acids).
Furthermore, we have determined the gene organization of
IHABP4 and cloned cDNAs for the chick, mouse, and human
homologs. Comparison of the deduced chick, mouse, and human protein
sequences showed that the hyaluronan-binding motifs,
(R/K)X7(R/K), in these sequences are
conserved; both chick and mouse IHABP4 were shown directly to bind
hyaluronan. Biochemical fractionation and immunofluorescent localization of epitope-tagged IHABP4 indicated that it is mainly present in the cytoplasm. These data support the possibility that intracellular hyaluronan and its binding proteins may play important roles in cell behavior.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF227683, AF227684, and AF241831.
Molecular Characterization of a Novel Intracellular
Hyaluronan-binding Protein*
*
This work was supported by National Institutes of Health
Grants CA73839 and CA82867 (to B. P. T.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 617-636-6659;
Fax: 617-636-0380; E-mail: bryan.toole@tufts.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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