Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
1999-03-17
2001-11-06
Carlson, Karen Cochrane (Department: 1653)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C530S350000, C435S069100, C435S069700, C435S320100, C435S252300, C435S069500, C435S325000, C536S023500
Reexamination Certificate
active
06313267
ABSTRACT:
BACKGROUND OF INVENTION
1. Field of the Invention
The present invention relates to novel calcium-binding proteins, DNA encoding them, antibodies against the calcium-binding proteins, hybridomas producing the antibodies, diagnostic agents comprising the antibodies, etc.
2. Related Art
Extracellular calcium ion concentration is known to play a role in the regulation of cellular proliferation and differentiation. On the other hand, intracellular calcium ion is one of the key transfer factors for intracellular signal transduction. The calcium signals are transduced by various calcium-binding proteins (CaBP). Calcium-binding proteins are largely classified as those with the EF-hand motif, such as calmodulin, troponin C, the S100 protein family, etc. and those without EF-hands, such as the anexine family. These are presumed to carry out separate and important physiological roles, but their physiological roles have not been completely elucidated.
Calmodulin mediates many calcium ion-dependent cellular reactions, and it is understood to be a ubiquitus CaBP required during mitosis. In contrast, CaBPs of the S100 protein family are expressed specifically according to cell cycle or cell type, and they are implicated in specific signal transduction for cellular division and differentiation. The S100 protein family includes S100&agr;, S100&bgr;, calcyclin, MRP8, MRP14, etc., each of which have 2 EF-hand motifs.
Thus, the study of the presence and function of CaBPs is crucial for understanding the mechanisms of cell proliferation and differentiation, and is expected to yield useful knowledge for the understanding, diagnosis and treatment of the related diseases.
SUMMARY OF INVENTION
The present invention, therefore, provides novel calcium-binding proteins, as well as methods for their production, gene systems for their production, antibodies against the proteins, and methods for use thereof.
The present inventors have searched for calcium-binding proteins from among various specimens, and have found that a protein with considerable calcium-binding activity is present in bovine amniotic fluid. In addition, the inventors have isolated the calcium-binding protein to obtain a substantially pure protein, and have determined its full amino acid sequence and the sequence of the DNA encoding it. This calcium-binding protein is a hitherto-unknown novel protein belonging to the S100 protein family and present in extracellular fluid, and it has been named CAAF1 (Calcium binding protein in Amniotic Fluid 1).
In addition the present inventors succeeded to clone a novel cDNA by screening a human cDNA library using as a probe the above-mentioned bovine cDNA. In addition, the present inventors determined a nucleotide sequence of the human cDNA and a deduced amino acid sequence of human calcium-binding protein. This human calcium-binding is novel as well.
Furthermore, the present inventors have prepared antibodies against CAAF1, have studied the presence of the proteins in various tissues, and have also constructed a system for a quantitative assay thereof and have found their usefulness as diagnostic agents for a number of diseases.
Thus, the present invention provides calcium-binding proteins which may comprise any amino acid sequence which is substantially identical to the amino acid sequence shown in SEQ ID NO: 1 or 12.
The present invention further provides DNAs encoding the above-mentioned proteins, expression vectors including the DNA, and host cells transformed with the expression vectors.
The present invention still further provides a method for producing the calcium-binding proteins by extraction from bovine amniotic fluid or tissue, human tissue, or by using the above-mentioned host cells.
The present invention still further provides antibodies against the calcium-binding proteins, hybridomas which produce the antibodies, and methods of producing the antibodies.
The present invention still further provides diagnostic agents comprising the antibodies and analysis methods employing them.
REFERENCES:
patent: 5288623 (1994-02-01), Zenno et al.
Dell'Angelica et al., The Journal of Biological Chemisty, vol. 269, No. 46, pp. 28929-28936, 1994.*
Guignard et al. Biochemical Journal, vol. 309, pp. 395-401, 1995.*
Alignments: Accession No. A55406, Dell'Angelica, 1994.
Hitomi Jiro
Kimura Tatsuji
Yamaguchi Ken
Yamamura Tokujiro
Advanced Life Science Institute, Inc.
Anderson Kill & Olick P.C.
Carlson Karen Cochrane
Robinson Hope A.
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