GSP-37, a novel goldfish scale matrix protein: Identification, localization and functional analysis

Kousei Miyabe, Hiroki Tokunaga, Hirotoshi Endo, Hirotaka Inoue, Michio Suzuki, Naoaki Tsutsui, Naoki Yokoo, Toshihiro Kogure, Hiromichi Nagasawa

Research output: Contribution to journalArticle

5 Citations (Scopus)

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 languageEnglish
Pages (from-to)463-481
Number of pages19
JournalFaraday Discussions
Volume159
DOIs
Publication statusPublished - 2012
Externally publishedYes

Fingerprint

functional analysis
Functional analysis
proteins
matrices
Proteins
Crystallization
phosphatases
Alkaline Phosphatase
crystallization
teeth
Protein Precursors
Enamels
Cell adhesion
Transcription
Protein Sorting Signals
cells
Phosphoric Monoester Hydrolases
Edetic Acid
Fish
Purification

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry

Cite this

Miyabe, K., Tokunaga, H., Endo, H., Inoue, H., Suzuki, M., Tsutsui, N., ... Nagasawa, H. (2012). GSP-37, a novel goldfish scale matrix protein: Identification, localization and functional analysis. Faraday Discussions, 159, 463-481. https://doi.org/10.1039/c2fd20051a

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 journalArticle

Miyabe, K, Tokunaga, H, Endo, H, Inoue, H, Suzuki, M, Tsutsui, N, Yokoo, N, Kogure, T & Nagasawa, H 2012, 'GSP-37, a novel goldfish scale matrix protein: Identification, localization and functional analysis', Faraday Discussions, vol. 159, pp. 463-481. https://doi.org/10.1039/c2fd20051a
Miyabe, Kousei ; Tokunaga, Hiroki ; Endo, Hirotoshi ; Inoue, Hirotaka ; Suzuki, Michio ; Tsutsui, Naoaki ; Yokoo, Naoki ; Kogure, Toshihiro ; Nagasawa, Hiromichi. / GSP-37, a novel goldfish scale matrix protein : Identification, localization and functional analysis. In: Faraday Discussions. 2012 ; Vol. 159. pp. 463-481.
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