Type XIII Collagen Is Identified as a Plasma Membrane Protein

• Published in: The Journal of biological chemistry 1998-06, Vol.273 (25), p.15590-15597
• Main Authors: Haegg, P, Rehn, M, Huhtala, P, Vaeisaenen, T, Tamminen, M, Pihlajaniemi, T
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Cell Line; Cell Membrane - chemistry; Cloning, Molecular; Collagen - chemistry; Collagen - genetics; DNA, Complementary - chemistry; Humans; Membrane Proteins - chemistry; Membrane Proteins - genetics; Mice; Molecular Sequence Data
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.25.15590

The complete primary structure of the mouse type XIII collagen chain was determined by cDNA cloning. Comparison of the mouse amino acid sequences with the previously determined human sequences revealed a high identity of 90%. Surprisingly, the mouse cDNAs extended further in the 5′ direction than the previously identified human clones. The 5′ sequences contained a new in-frame ATG codon for translation initiation which resulted in elongation of the N-terminal noncollagenous domain by 81 residues. These N-terminal sequences lack a typical signal sequence but include a highly hydrophobic segment that clearly fulfills the criteria for a transmembrane domain. The sequence data thus unexpectedly suggested that type XIII collagen may be located on the plasma membrane, with a short cytosolic N-terminal portion and a long collagenous extracellular portion. These sequence data prompted us to generate antipeptide antibodies against type XIII collagen in order to study the protein and its subcellular location. Western blotting of human tumor HT-1080 cell extract revealed bands of over 180 kDa. These appeared to represent disulfide-bonded multimeric polypeptide forms that resolved upon reduction into 85–95-kDa bands that are likely to represent a mixture of splice forms of monomeric type XIII collagen chains. These chains were shown to contain the predicted N-terminal extension and thus also the putative transmembrane segment. Immunoprecipitation of biotinylated type XIII collagen from surface-labeled HT-1080 cells, subcellular fractionation, and immunofluorescence staining were used to demonstrate that type XIII collagen molecules are indeed located in the plasma membranes of these cells.