Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1996-05-30
1999-06-01
Hutzell, Paula K.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
536 231, 4353201, 4352523, 5303871, 5303879, 530350, C07H 1900, C12P 2106, C07K 100, C07K 1600
Patent
active
059087617
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention is generally in the field of mammalian S-type lectin proteins, now designated galectins, which are thiol-dependent and specifically bind .beta.-galactoside residues.
More specifically, the present invention relates to a new S-type mammalian lectin, termed hereinafter as "galectin-8", and to galectin-8-like proteins, to DNA molecules coding therefor and to antibodies raised against said proteins. The invention further relates to pharmaceutical compositions comprising said proteins for the purpose of cell growth regulation in general, and more particularly for inhibition of cell proliferation and for treatment of tumors.
BACKGROUND OF THE INVENTION
Lectins are involved in a wide variety of cellular functions, many of which are related to their only common feature, the ability to bind carbohydrates specifically and reversibly, and to agglutinate cells Ca.sup.2+ -dependent and are structurally related to the asialoglycoprotein receptor, and galectins, previously known as S-type lectins, which are thiol-dependent and specifically bind .beta.-galactoside residues. In mammals, four galectin types have been sequenced and characterized, and there is evidence for the existence of other relatives (2,3). All known members of this family lack a signal peptide, are found in the cytosol, and are isolated as soluble proteins. However, there is evidence that some members are externalized by an atypical secretory mechanism.
Galectins require fulfillment of two criteria: affinity for .beta.-galactosides and significant sequence similarity in the carbohydrate recognition domain (CRD) (4), the relevant amino acids residues of which have been determined by X-ray crystallography (5). Galectin-1 and -2 are homodimers with subunit molecular weight of .apprxeq.14 kDa, that are not subjected to post-translational modifications (6). Galectin-1 is found in the extracellular matrix and has been shown to interact with laminin (7). The function of galectin-1 and -2 is not yet fully understood, although there is evidence that they might be involved in regulation of cell growth (8); cell adhesion (7); cell transformation (9); and embryogenesis (10).
Larger galectins (galectin-3) (previously known as CBP-35, Mac-2, RL-29) do exist ((11) and references therein). These are monomeric 29-35 kDa mosaic proteins, composed of an N-terminal half made of tandem repeats characteristic of the collagen gene superfamily, and a C-terminal half homologous to galectins-1 and -2 (11). Galectin-3 also binds laminin, and is implicated as component of growth regulatory systems; mediator of cell--cell and cell-matrix interactions; modulator of immune response; marker of neoplastic transformation, and indicator for metastatic potential of melanoma cells.
Galectin-4 was cloned from rat intestine (12), and an homologous protein was cloned from nematode (13). Galectin-4 is a monomer with molecular mass of 36 kDa. It contains tandem domains of .apprxeq.140 amino-acids each, homologous to galectin-1 and -2, that are separated by a link region (12). The function of galectin-4 is presently unknown.
Galectins may functionally substitute each other. The absence of any major phenotypic abnormalities in mice carrying a null mutation in the gene encoding galectin-1, suggests that other protein(s), presumably galectin-3, are capable of functionally substituting for galectin-1, at least at early stages of embryogenesis.
It is an object of the present invention to provide the cloning of a cDNA encoding for a novel protein that we term galectin-8. Galectin-8 has the characteristic properties of other galectins (2,3), and it is structurally related (34% identity) to rat galectin-4 (12).
SUMMARY OF THE INVENTION
According to the present invention, a novel protein of 35 Kd which has the characteristic properties of galectins (S-type mammalian lectins) was cloned from a rat liver cDNA expression library. This protein was originally called by us RL-30 protein. However, the nomenclature of S-type lectins has recently been changed to
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Hutzell Paula K.
Ungar Susan
Yeda Research and Development Co. Ltd.
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