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Analysis of Proteinaceous Components of the Organic Matrix of Endoskeletal Sclerites from the Alcyonarian Lobophytum crassum

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Abstract

The mesoglea of alcyonarians is occupied by an abundance of minute calcitic sclerites. The sclerites of the alcyonarian Lobophytum crassum contain a water-soluble organic matrix comprising 0.48% of the sclerite weight and a water-insoluble fraction comprising 1.15% of the sclerite weight. Analysis of proteinaceous components in the soluble fraction shows a particularly high content of aspartic acid, followed by alanine, glycine, and glutamate. Aspartic acid, glycine, alanine, and glutamate are the most abundant residues in the insoluble fraction. In both cases, the fractions show the highest concentration of aspartic acid from the total proteins. In an in vitro assay, we show that the matrix proteins extracted from the calcitic sclerites induce the formation of amorphous calcium carbonate prior to its transformation into the calcitic crystalline form. We also show scanning electron micrographs of the rhombohedral calcite crystals used as template, the protein imprinted with these crystals. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of both matrices shows the protein fractions at 67 and 48 kDa. The soluble matrix shows two additional faint bands. Both fractions stain for a carbohydrate at 67 kDa, indicating a glycoprotein at this molecular weight. A newly derived protein sequence was subjected to bioinformatics analysis involving identification of similarities to other acidic proteins. The identification of these proteins in alcyonarian endoskeletal sclerites emphasizes the fundamental importance of such acidic proteins and sheds more light on the functions of these proteins in the processes of biocalcification.

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Acknowledgments

This study was supported by a grant from the 21st Century COE project “The Comprehensive Analyses on Biodiversity in Coral Reef and Island Ecosystems in Asian and Pacific Regions” from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (Monbukagakusho). The first author is grateful to the Rotary Yoneyama Memorial Foundation, Japan, for scholarship grants and especially to all the members of Ginowan Rotary Club, Okinawa, for their great help and cooperation during the period of the grants.

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Authors and Affiliations

  1. Department of Marine and Environmental Sciences, Faculty of Science, University of the Ryukyus, 1 Senbaru, Nishihara-cho, Okinawa, 903-0213, Japan

    M. Azizur Rahman, Y. Isa & T. Uehara

  2. Tropical Biosphere Research Center, University of the Ryukyus, 3422 Sesoko, Motobu, Okinawa, 905-0227, Japan

    A. Takemura

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  1. M. Azizur Rahman
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  2. Y. Isa
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  3. A. Takemura
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  4. T. Uehara
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Correspondence to M. Azizur Rahman.

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Rahman, M.A., Isa, Y., Takemura, A. et al. Analysis of Proteinaceous Components of the Organic Matrix of Endoskeletal Sclerites from the Alcyonarian Lobophytum crassum. Calcif Tissue Int 78, 178–185 (2006). https://doi.org/10.1007/s00223-005-0253-y

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  • DOI: https://doi.org/10.1007/s00223-005-0253-y

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