Cloning and Deduced Amino Acid Sequence of a Novel Cartilage Protein (CILP) Identifies a Proform Including a Nucleotide Pyrophosphohydrolase

The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. A single 4.2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a cal...

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Bibliographic Details
Published in:The Journal of biological chemistry 1998-09, Vol.273 (36), p.23469-23475
Main Authors: Lorenzo, Pilar, Neame, Peter, Sommarin, Yngve, Heinegård, Dick
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Cartilage - chemistry
Chondrocytes - metabolism
Cloning, Molecular
COS Cells
DNA, Complementary - genetics
Extracellular Matrix Proteins - chemistry
Extracellular Matrix Proteins - genetics
Extracellular Matrix Proteins - metabolism
Glycoproteins - chemistry
Glycoproteins - genetics
Glycoproteins - metabolism
Humans
Molecular Sequence Data
Peptide Mapping
Polymerase Chain Reaction
Protein Precursors - genetics
Protein Processing, Post-Translational
Pyrophosphatases - genetics
Recombinant Proteins - biosynthesis
Restriction Mapping
Sequence Analysis
Sequence Homology, Amino Acid
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Summary:The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. A single 4.2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). CILP has 30 cysteines and six putative N-glycosylation sites. The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.273.36.23469