Abstract
A novel noncollagenous acidic protein was identified from the scales of goldfish (Carassius auratus), a freshwater teleost. Using an in vitro calcium phosphate crystallization assay, the EDTA-soluble fraction from these scales was screened for crystallization inhibitory activity, and a highly phosphorylated glycoprotein, named goldfish scale protein (GSP)-37, was isolated through 5 HPLC purification steps. The cDNA for GSP-37 has an open reading frame encoding a precursor protein, consisting of a signal peptide and GSP-37, with 19 and 137 amino acid residues, respectively. The C-terminal region of GSP-37 contains the RGD consensus sequence for cell adhesion. Although native GSP-37 strongly inhibited crystallization, alkaline phosphatase treatment dramatically reduced its inhibitory activity. Reverse transcription-PCR analysis revealed that GSP-37 is expressed only in scales but not in other calcified tissues, bones or pharyngeal teeth. In situ hybridization demonstrated that GSP-37-expressing cells were localized in the central regions of regenerating scales, where organic matrices were actively synthesized and were not stained with either alkaline phosphatase or tartrate-resistant acidic phosphatase, osteoblastic and osteoclastic cell markers, respectively. Immunohistochemical analyses showed that GSP-37 is localized in the uppermost region of the bony layer of the scale, which is thought to correspond to the enamel or enameloid layer of vertebrate teeth. All these data strongly indicate that GSP-37 is deeply associated with calcification in fish scales.
| Original language | English |
|---|---|
| Pages (from-to) | 463-481 |
| Number of pages | 19 |
| Journal | Faraday Discussions |
| Volume | 159 |
| DOIs | |
| Publication status | Published - 2012 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Physical and Theoretical Chemistry
Cite this
GSP-37, a novel goldfish scale matrix protein : Identification, localization and functional analysis. / Miyabe, Kousei; Tokunaga, Hiroki; Endo, Hirotoshi; Inoue, Hirotaka; Suzuki, Michio; Tsutsui, Naoaki; Yokoo, Naoki; Kogure, Toshihiro; Nagasawa, Hiromichi.
In: Faraday Discussions, Vol. 159, 2012, p. 463-481.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - GSP-37, a novel goldfish scale matrix protein
T2 - Identification, localization and functional analysis
AU - Miyabe, Kousei
AU - Tokunaga, Hiroki
AU - Endo, Hirotoshi
AU - Inoue, Hirotaka
AU - Suzuki, Michio
AU - Tsutsui, Naoaki
AU - Yokoo, Naoki
AU - Kogure, Toshihiro
AU - Nagasawa, Hiromichi
PY - 2012
Y1 - 2012
N2 - A novel noncollagenous acidic protein was identified from the scales of goldfish (Carassius auratus), a freshwater teleost. Using an in vitro calcium phosphate crystallization assay, the EDTA-soluble fraction from these scales was screened for crystallization inhibitory activity, and a highly phosphorylated glycoprotein, named goldfish scale protein (GSP)-37, was isolated through 5 HPLC purification steps. The cDNA for GSP-37 has an open reading frame encoding a precursor protein, consisting of a signal peptide and GSP-37, with 19 and 137 amino acid residues, respectively. The C-terminal region of GSP-37 contains the RGD consensus sequence for cell adhesion. Although native GSP-37 strongly inhibited crystallization, alkaline phosphatase treatment dramatically reduced its inhibitory activity. Reverse transcription-PCR analysis revealed that GSP-37 is expressed only in scales but not in other calcified tissues, bones or pharyngeal teeth. In situ hybridization demonstrated that GSP-37-expressing cells were localized in the central regions of regenerating scales, where organic matrices were actively synthesized and were not stained with either alkaline phosphatase or tartrate-resistant acidic phosphatase, osteoblastic and osteoclastic cell markers, respectively. Immunohistochemical analyses showed that GSP-37 is localized in the uppermost region of the bony layer of the scale, which is thought to correspond to the enamel or enameloid layer of vertebrate teeth. All these data strongly indicate that GSP-37 is deeply associated with calcification in fish scales.
AB - A novel noncollagenous acidic protein was identified from the scales of goldfish (Carassius auratus), a freshwater teleost. Using an in vitro calcium phosphate crystallization assay, the EDTA-soluble fraction from these scales was screened for crystallization inhibitory activity, and a highly phosphorylated glycoprotein, named goldfish scale protein (GSP)-37, was isolated through 5 HPLC purification steps. The cDNA for GSP-37 has an open reading frame encoding a precursor protein, consisting of a signal peptide and GSP-37, with 19 and 137 amino acid residues, respectively. The C-terminal region of GSP-37 contains the RGD consensus sequence for cell adhesion. Although native GSP-37 strongly inhibited crystallization, alkaline phosphatase treatment dramatically reduced its inhibitory activity. Reverse transcription-PCR analysis revealed that GSP-37 is expressed only in scales but not in other calcified tissues, bones or pharyngeal teeth. In situ hybridization demonstrated that GSP-37-expressing cells were localized in the central regions of regenerating scales, where organic matrices were actively synthesized and were not stained with either alkaline phosphatase or tartrate-resistant acidic phosphatase, osteoblastic and osteoclastic cell markers, respectively. Immunohistochemical analyses showed that GSP-37 is localized in the uppermost region of the bony layer of the scale, which is thought to correspond to the enamel or enameloid layer of vertebrate teeth. All these data strongly indicate that GSP-37 is deeply associated with calcification in fish scales.
UR - http://www.scopus.com/inward/record.url?scp=84867947997&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84867947997&partnerID=8YFLogxK
U2 - 10.1039/c2fd20051a
DO - 10.1039/c2fd20051a
M3 - Article
AN - SCOPUS:84867947997
VL - 159
SP - 463
EP - 481
JO - Faraday Discussions
JF - Faraday Discussions
SN - 1359-6640
ER -