Abstract
Two species of alcyonarian corals, Lobophytum crassum and Sinularia polydactyla, are closely related to each other. It is reported that the calcified organic substances in the skeletons of both contain a protein–polysaccharide complex playing a key role in the regulation of biocalcification. However, information on the matrix proteins of endoskeletal sclerite has been lacking. Hence we studied the proteinaceous organic matrices of sclerites for both species, to analyze the sequences and the functional properties of the proteins present. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the preparations showed four bands of proteins with apparent molecular masses of 102, 67, 48, and 37 kDa for L. crassum and seven bands of 109, 83, 70, 63, 41, 30, and 22 kDa for S. polydactyla. A major protein band of about 67 kDa in L. crassum and two bands of proteins of about 70 and 63 kDa in S. polydactyla yielded N-terminal amino acid sequences. Periodic acid-Schiff staining indicated that the 67-kDa protein in L. crassum, and 83- and 63-kDa proteins in S. polydactyla were glycosylated. For detection of calcium binding proteins, a Ca2+ overlay analysis was conducted in the extract via 45Ca autoradiography. The 102- and 67-kDa calcium binding proteins in L. crassum, and the 109- and 63-kDa Ca2+ binding proteins in S. polydactyla were found to be radioactive. An assay for carbonic anhydrase (CA), which is thought to play an important role in the process of calcification, revealed specific activities. Newly derived protein sequences were subjected to standard sequence analysis involving identification of similarities to other proteins in databases. The significantly different protein expressions and compositional analysis of sequences between two species were demonstrated.
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References
HH Boer J Witteveen (1980) ArticleTitleUltrastructural localization of carbonic anhydrase in tissues involved in shell formation and ionic regulation in the pond snail Lymnaea stagnalis Calcif Tissue Res 209 383–390
V Brendel P Bucher I Nourbakhsh BE Blaisdell S Karlin (1992) ArticleTitleMethods and algorithms for statistical analysis of protein sequences Proc Natl Acad Sci USA 89 2002–2006 Occurrence Handle10.1073/pnas.89.6.2002
JW Cho JS Partin WJ Lennarz (1996) ArticleTitleA technique for detecting matrix proteins in the crystalline spicule of the sea urchin embryo Proc Natl Acad Sci USA 93 1282–1286 Occurrence Handle10.1073/pnas.93.3.1282
MM Giraud (1981) ArticleTitleCarbonic anhydrase activity in the integument of the crab Carcinus maenas during the intermolt cycle Comp Biochem Physiol 69A 381–387 Occurrence Handle10.1016/0300-9629(81)92993-5
BM Heatfield (1970) ArticleTitleCalcification in echinoderms: effects of temperature and Diamox on incorporation of calcium-45 in vitro by regenerating spines of Strongylocentrotus purpurratus Biol Bull 139 151–163
C Jolly S Berland C Milet S Borzeix E Lopez D Doumenc (2004) ArticleTitleZonal localization of shell matrix proteins in mantle of Haliots tuberculata (Mollusca, Gastropoda) Mar Biotechnol 6 541–551 Occurrence Handle10.1007/s10126-004-3129-7
RJ Kingsley N Watabe (1983) ArticleTitleAnalysis of proteinaceous components of the organic matrices of spicules from the gorgonian Leptogorgia virgulata Comp Biochem Physiol 76B 443–447
UK Laemmli (1970) ArticleTitleCleavage of structural proteins during assembly of the head of bacteriophage T4 Nature 227 680–685 Occurrence Handle10.1038/227680a0
OH Lowry NJ Rosebrough AL Farr RJ Randall (1951) ArticleTitleProtein measurement with the Folin phenol reagent J Biol Chem 193 265–275
TH Maren (1960) ArticleTitleA simplified micromethod for the determination of carbonic anhydrates and its inhibitors J Pharmacol Exp Ther 130 26–29
TH Maren (1967) ArticleTitleCarbonic anhydrase: chemistry, physiology and inhibition Physiol Rev 47 595–781
K Maruyama T Mikawa S Ebashi (1984) ArticleTitleDetection of calcium binding proteins by 45Ca autoradiography on nitrocellulose membrane after sodium dodecyl sulphate gel electrophoresis J Biochem 95 511–519
A Matsushiro T Miyashita H Miyamoto K Moromoto B Tonomura A Tanaka K Sato (2003) ArticleTitlePresence of protein complex is prerequisite for aragonite crystallization in the nacreous layer Mar Biotechnol 5 37–44 Occurrence Handle10.1007/s10126-002-0048-3
RM Mitterer (1978) ArticleTitleAmino acid composition and metal binding capability of the skeletal protein of corals Bull Mar Sci 28 173–180
T Miyashita Y Takagi M Okushima S Nakano H Miyamoto E Nishikawa A Matsushiro (2000) ArticleTitleComplementary DNA cloning and characterization of pearlin, a new class of matrix protein in the nacreous layer of oyster pearls Mar Biotechnol 2 120–129
MA Rahman Y Isa T Uehara (2005) ArticleTitleProteins of calcified endoskeleton: II. Partial amino acid sequences of endoskeletal proteins and the characterization of proteinaceous organic matrix of spicules from the alcyonarian, Synularia polydactyla Proteomics 5 885–893 Occurrence Handle10.1002/pmic.200401130
WJ Schmidt (1924) Die Bausteine des Tierkorpers in Polarisiertem Lichte F. Cohen Verlag Bonn
Segrest JP, Jackson RL (1972) Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. In: Ginsberg V, ed., Methods in Enzymology, Vol. 28 ( New York Academic Press) pp 54–63
K Simkiss K Wilbur (1989) Biomineralization, Cell Biology and Mineral Deposition Academic Press New York
TF Smith MS Waterman (1981) ArticleTitleIdentification of common molecular subsequences J Mol Biol 147 195–197 Occurrence Handle10.1016/0022-2836(81)90087-5
JD Thompson F Plewnial J–C Thierry O Poch (2000) ArticleTitleRapid and reliable global multiple alignments of protein sequences detected by database searches Nucleic Acids Res 28 IssueID15 2919–2926 Occurrence Handle10.1093/nar/28.15.2919
AS Tompa KM Wilbur JH Waite (1977) ArticleTitleStructural proteins in the calcified egg shell of the giant land snail Strophocheilus oblongus (Becquaert) Comp Biochem Physiol 56B 279–283
S Weiner (1979) ArticleTitleAspartic acid-rich proteins: major components of the soluble organic matrix of mollusk shells Calcif Tissue Int 29 163–167 Occurrence Handle10.1007/BF02408072
S Weiner L Hood (1975) ArticleTitleSoluble protein of the organic matrix of mollusk shells: a potential template for shell formation Science 190 987–989
Weiner SW, Traub W, Lowenstam HA (1983) Organic matrix in calcified exoskeletons. In: Westbroek P, de Jong EW, eds., Biomin. and Biol Met Accum, Reidel, Dordrecht, pp 205–224
AP Wheeler (1981) ArticleTitleControl of calcium carbonate nucleation and crystal growth by soluble matrix of oyster shell Science 212 1397–1398
M Wilfert W Peters (1969) ArticleTitleVorkommon von Chitin bei Coelenteraten Z Morphol Tiere 64 77–84 Occurrence Handle10.1007/BF00300386
S–Y Yang M Tsuzuki S Miyachi (1985) ArticleTitleCarbonic anhydrase of Chlamydomonas: Purification and studies on its induction using antiserum against Chlamydomonas carbonic anhydrase Plant Cell Physiol 26 25–34
SD Young (1971) ArticleTitleOrganic material from scleractinian coral skeletons-I. Variation in composition between several species Comp Biochem Physiol 40B 113–120
Acknowledgments
This study was partially supported by the 21st Century COE program of the University of the Ryukyus. M. Azizur Rahman 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 grant period.
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Rahman, M.A., Isa, Y. & Uehara, T. Studies on Two Closely Related Species of Octocorallians: Biochemical and Molecular Characteristics of the Organic Matrices of Endoskeletal Sclerites. Mar Biotechnol 8, 415–424 (2006). https://doi.org/10.1007/s10126-005-6150-6
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DOI: https://doi.org/10.1007/s10126-005-6150-6