Identification of proteoglycan from salmon nasal cartilage.
There has been no structural information about the core protein of salmon nasal cartilage proteoglycan although its physiological activities have been investigated. Internal amino acid sequencing using nano-LC/MS/MS revealed that the salmon proteoglycan was aggrecan. Primer walk sequencing based on the amino acid information determined that the salmon aggrecan cDNA is comprised of 4207bp nucleotides predicted to encode 1324 amino acids with a molecular mass of 143,276. It exhibited significant similarities to predicted pufferfish aggrecan, zebrafish similar to aggrecan, zebrafish aggrecan, bovine aggrecan and human aggrecan isoform 2 precursor; whose amino acid identities were 56%, 55%, 49%, 31% and 30%, respectively. Salmon cartilage aggrecan had globular domains G1, G2 and G3 as in mammalian aggrecans. Neither the putative keratan sulfate attachment domain enriched with serine, glutamic acid and proline, nor the putative chondroitin sulfate attachment domain with repeating amino acid sequence containing serine-glycine, found in mammalian aggrecans were observed in salmon, however, random serine-glycine (or glycine-serine) sequences predicted to the sugar chain attachment sites were observed. Based on cDNA analysis and amino acid analysis after β-elimination, the ratio of serine attached to sugar chains was calculated to be approximately 37.7% of total serine, that is, 46 of 123 serine residues.
Department of Glycotechnology, Center for Advanced Medical Research, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki 036-8562, Japan. kaki@cc.hirosaki-u.ac.jp
Ikuko
I
Kakizaki
Yota
Y
Tatara
Mitsuo
M
Majima
Yoji
Y
Kato
Masahiko
M
Endo
D053674:Aggrecans / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D029941:Fish Proteins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D055171:Nasal Cartilages / Q000737:chemistry*; D017685:Oncorhynchus keta / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000737:chemistry* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D053674:Aggrecans; D000596:Amino Acids; D017931:DNA Primers; D018076:DNA, Complementary; D029941:Fish Proteins; D011509:Proteoglycans