地中海贻贝贝壳肌棱柱层蛋白质组学分析
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摘要
贻贝贝壳与后闭壳肌形成紧密连接并由此介导了贝壳的闭合。为了解贻贝后闭壳肌-贝壳之间连接界面以及蛋白质分子组成,利用扫描电子显微镜、傅里叶红外光谱对地中海贻贝贝壳的微观结构、碳酸钙晶体构型进行分析;同时,采用酸抽提法获取地中海贻贝后闭壳肌痕部位肌棱柱层的贝壳基质蛋白,对酸可溶性蛋白和酸不溶性蛋白分别采用液质联用技术结合转录组数据库搜索,共鉴定61种贝壳基质蛋白。上述研究为深入了解贝壳的生物矿化机制以及肌肉-贝壳界面的无机相-有机相连接机理奠定了基础。
The attachment of adductor muscle-shell in Mytilus mediates the closing of shells. For understanding the mechanism and the molecular composition of this attachment, Scanning Electronic Microscopic and Fourier Transform Infrared Spectroscopy were used to explore the micro-structure and polymorph of muscle-shell attachment of Mytilus galloprovincialis. Furthermore, the protein composition of shell matrix from myostracum layer from adductor muscle scar of M. galloprovincialis was detected by a combination of LC-MS/MS analysis with the Mytilus EST dataset search, which resulted in the identification of a total of 61 proteins from acid-soluble and acid-insoluble shell matrix proteins form myostracum of M. galloprovincialis shell. From this protein set, many novel shell proteins were identified which included proteins with possible link to biomin-eralization and certain uncharacterized proteins with unusual amino acid composition. This data would be use-ful in understanding the role of SMPs associated with the formation of myostracum. Further, the identified pro-tein set from the myostracum layer could provide a clue for exploring the mechanism of adductor muscle-shell attachment.
The attachment of adductor muscle-shell in Mytilus mediates the closing of shells. For understanding the mechanism and the molecular composition of this attachment, Scanning Electronic Microscopic and Fourier Transform Infrared Spectroscopy were used to explore the micro-structure and polymorph of muscle-shell attachment of Mytilus galloprovincialis. Furthermore, the protein composition of shell matrix from myostracum layer from adductor muscle scar of M. galloprovincialis was detected by a combination of LC-MS/MS analysis with the Mytilus EST dataset search, which resulted in the identification of a total of 61 proteins from acid-soluble and acid-insoluble shell matrix proteins form myostracum of M. galloprovincialis shell. From this protein set, many novel shell proteins were identified which included proteins with possible link to biomin-eralization and certain uncharacterized proteins with unusual amino acid composition. This data would be use-ful in understanding the role of SMPs associated with the formation of myostracum. Further, the identified pro-tein set from the myostracum layer could provide a clue for exploring the mechanism of adductor muscle-shell attachment.
引文
[1]ALBECK S,ADDADI L,WEINER S.Regulation of calcite crystal morphology by intracrystalline acidic proteins and glycoproteins[J].Connective Tissue Research,1996,35:365-370.
[2]SELLINGER A,WEISS P M,NGUYEN A,et al.Continuous selfassembly of organic-inorganic nanocomposite coatings that mimic nacre[J].Nature,1998,394:256-260.
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[5]JACKSON A P,VINCENT J F V,TURNER R M.The mechanical design of nacre[J].Proc R Soc Lond B Biol Sci,1988,234:415-440.
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[7]LI X,CHANG W C,YUH J C,et al.Nanoscale structural and mechanical characterization of a natural nanocomposite material:The shell of red abalone[J].Nano Lett,2004,4:613-617.
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[13]SUZUKI M,SAKUDA S,NAGASAWA H.Identification of chitin in the prismatic layer of the shell and a chitin synthase gene from the Japanese pearl oyster,Pinctada fucata[J].Bioscience,biotechnology,and biochemistry,2007,71:1 735-1 744.
[14]MARIE B,JOUBERT C,TAYAL A,et al.Different secretory repertoires control the biomineralization processes of prism and nacre deposition of the pearl oyster shell[J].Proc Natl Acad Sci,2012,109(51):20 986-20 991.
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[17]CAROLINE J,DAVID P,BENJAMIN M.Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell:focus on biomineralization[J].BMC Genomics,2010,11:613-625.
[18]MARIE B,MARIE A,JACKSON D J.Proteomic analysis of the organic matrix of the abalone Haliotis asinina calcified shell[J].Proteome Science,2010,8:54-64.
[19]MARIE B,TRINKLER N,ZANELLA-CLEON I.Proteomic identification of novel proteins from the calcifying shell matrix of the Manila clam Venerupis philippinarum[J].Marine Biotechnology,2011,13:955-962.
[20]LIN J Y,MA K Y,BAI Z Y,et al.Molecular cloning and characterization of perlucin from the freshwater pearl mussel,Hyriopsis cumingii[J].Gene,2013,526(2):210-216.
[21]MIYASHITA T,TAKAGI R,MIYAMOTA H,et al.Identical carbonic anhydrase contributes to nacreous or prismatic layer formation in Pinctada fucata(Mollusca:Bivalvia)[J].Veliger,2002,45:250-255.
[22]MARIE B,MARIN F,MARIE A,et al.Evolution of nacre:Biochemistry and proteomics of the shell organic matrix of the cephalopod Nautilus macromphalus[J].Chembiochem,2009,10:1 495-1 506.
[23]MIYAMOTO H,YANO M,MIYASHITA T.Similarities in the structure of nacrein,the shell-matrix protein,in a bivalve and a gastropod[J].J Molluscan Stud,2003,69:87-89.
[24]JACKSON D J,MCDOUGALL C,WOODCROFT B,et al.Parallel evolution of nacre building gene sets in molluscs[J].Molecular Biology Evolution,2010,27:591-608.
[25]ZHANG G,FANG X,GUO X,et al.The oyster genome reveals stress adaptation and complexity of shell formation[J].Nature,2012,490:49-54.
[26]KHAN Z,LAFON M.PDZ domain-mediated protein interactions:therapeutic targets in neurological disorder s[J].Curr Med Chem,2014,21(23):2 632-2 641.
[27]CHI C N,BACH A,STR?MGAARD K,et al.Ligand binding by PDZ domains[J].Biofactors,2012,38(5):338-348.
[28]FERJANI I,FATTOUM A,MANAI M,et al.Two distinct regions of calponin share common binding sites on actin resulting in different modes of calponin-actin interaction[J].Biochimica et Biophysica Acta,2010,1804:1 760-1 767.
[29]XUE B,LEYRAT C,GRIMES J M,et al.Structural basis of thymosin-β4/profilin exchange leading to actin filament polymerization[J].Proc Natl Acad Sci,2014,111(43):4 596-4 605.
[30]RAMAN M,MATHEW S.Study of chemical properties and evaluation of collagen in mantle,epidermal connective tissue and tentacle of Indian Squid,Loligo duvauceli Orbigny[J].J Food Sci Technol,2014,51(8):1509-16.
[31]COLOMBATTI A,BONALDO P,DOLIANA R.Type A modules:interacting domains found in several non-fibrillar collagens and in other extracellular matrix proteins[J].Matrix,1993,13(4):297-306.
[32]RUSKAMO S,YLNNE J.Structure of the human filamin A actin-binding domain[J].Acta Crystallogr D Biol Crystallogr,2009,65(11):1 217-1 221.
[33]HERRMANN H,B?R H,KREPLAK L,et al.Intermediate filaments:from cell architecture to nanomechanics[J].Nat Rev Mol Cell Biol,2007,8(7):562-573.
[34]EVANS J S.“Tuning in”to mollusk shell nacre-and prismatic-associated protein terminal sequence.Implications for biomineralization and the construction of high performance inorganic-organic composites[J].Chemical Reviews,2008,108:4 455-4462.
[2]SELLINGER A,WEISS P M,NGUYEN A,et al.Continuous selfassembly of organic-inorganic nanocomposite coatings that mimic nacre[J].Nature,1998,394:256-260.
[3]LEE S W,JANG Y N,RYU K W,et al.Mechanical characteristics and morphological effect of complex crossed structure in biomaterials:fracture mechanics and microstructure of chalky layer in oyster shell[J].Micron,2011,42:60-70.
[4]KOUCHINSKY A.Shell microstructures in Early Cambrian mollusks[J].Acta Palaeontol Pol,2000,45(2):119-150.
[5]JACKSON A P,VINCENT J F V,TURNER R M.The mechanical design of nacre[J].Proc R Soc Lond B Biol Sci,1988,234:415-440.
[6]TAYLOR J,LAYMAN M.The mechanical properties of bivalve(Mollusca)shell structures[J].Paleontology,1972,15(1):73-87.
[7]LI X,CHANG W C,YUH J C,et al.Nanoscale structural and mechanical characterization of a natural nanocomposite material:The shell of red abalone[J].Nano Lett,2004,4:613-617.
[8]LEE S W,JANG Y N,KIM J C.Characteristics of the Aragonitic Layer in Adult Oyster Shells,Crassostrea gigas:Structural Study of Myostracum including the Adductor Muscle Scar[J].Evid Based Complement Alternat Med,2011,2011:742 963.
[9]LOWENSTAM H A,WEINER S.On Biomineralization[M].New York:Oxford University Press,1989.
[10]CARTER J G.Shell microstructural data for the Bivalvia[Z]//Skeletal Biomineralization:Patterns,Processes and Evolutionary Trends.New York:Van Nostrand Reinhold,1990,1:297-411.
[11]GALTSOFF P A.The American oyster.Fish and Wildlife Service[M].U.S.Government Printing Office,1964.
[12]ALBECK S,ADDADI L,WEINER S.Regulation of calcite crystal morphology by intracrystalline acidic proteins and glycoproteins[J].Connective Tissue Research,1996,35:365-370.
[13]SUZUKI M,SAKUDA S,NAGASAWA H.Identification of chitin in the prismatic layer of the shell and a chitin synthase gene from the Japanese pearl oyster,Pinctada fucata[J].Bioscience,biotechnology,and biochemistry,2007,71:1 735-1 744.
[14]MARIE B,JOUBERT C,TAYAL A,et al.Different secretory repertoires control the biomineralization processes of prism and nacre deposition of the pearl oyster shell[J].Proc Natl Acad Sci,2012,109(51):20 986-20 991.
[15]DAUPHIN Y.Infrared spectra and elemental composition in recent biogenic calcites:relationships between theυ4 band wavelength and Sr and Mg concentrations[J].Appl Spectrom,1999,53:184-190.
[16]MARIE B,LE RN,ZANELLA-CLEON I,et al.Molecular evolution of mollusc shell proteins:insights from proteomic analysis of the edible mussel Mytilus[J].J Mol Evol,2011,72:531-546.
[17]CAROLINE J,DAVID P,BENJAMIN M.Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell:focus on biomineralization[J].BMC Genomics,2010,11:613-625.
[18]MARIE B,MARIE A,JACKSON D J.Proteomic analysis of the organic matrix of the abalone Haliotis asinina calcified shell[J].Proteome Science,2010,8:54-64.
[19]MARIE B,TRINKLER N,ZANELLA-CLEON I.Proteomic identification of novel proteins from the calcifying shell matrix of the Manila clam Venerupis philippinarum[J].Marine Biotechnology,2011,13:955-962.
[20]LIN J Y,MA K Y,BAI Z Y,et al.Molecular cloning and characterization of perlucin from the freshwater pearl mussel,Hyriopsis cumingii[J].Gene,2013,526(2):210-216.
[21]MIYASHITA T,TAKAGI R,MIYAMOTA H,et al.Identical carbonic anhydrase contributes to nacreous or prismatic layer formation in Pinctada fucata(Mollusca:Bivalvia)[J].Veliger,2002,45:250-255.
[22]MARIE B,MARIN F,MARIE A,et al.Evolution of nacre:Biochemistry and proteomics of the shell organic matrix of the cephalopod Nautilus macromphalus[J].Chembiochem,2009,10:1 495-1 506.
[23]MIYAMOTO H,YANO M,MIYASHITA T.Similarities in the structure of nacrein,the shell-matrix protein,in a bivalve and a gastropod[J].J Molluscan Stud,2003,69:87-89.
[24]JACKSON D J,MCDOUGALL C,WOODCROFT B,et al.Parallel evolution of nacre building gene sets in molluscs[J].Molecular Biology Evolution,2010,27:591-608.
[25]ZHANG G,FANG X,GUO X,et al.The oyster genome reveals stress adaptation and complexity of shell formation[J].Nature,2012,490:49-54.
[26]KHAN Z,LAFON M.PDZ domain-mediated protein interactions:therapeutic targets in neurological disorder s[J].Curr Med Chem,2014,21(23):2 632-2 641.
[27]CHI C N,BACH A,STR?MGAARD K,et al.Ligand binding by PDZ domains[J].Biofactors,2012,38(5):338-348.
[28]FERJANI I,FATTOUM A,MANAI M,et al.Two distinct regions of calponin share common binding sites on actin resulting in different modes of calponin-actin interaction[J].Biochimica et Biophysica Acta,2010,1804:1 760-1 767.
[29]XUE B,LEYRAT C,GRIMES J M,et al.Structural basis of thymosin-β4/profilin exchange leading to actin filament polymerization[J].Proc Natl Acad Sci,2014,111(43):4 596-4 605.
[30]RAMAN M,MATHEW S.Study of chemical properties and evaluation of collagen in mantle,epidermal connective tissue and tentacle of Indian Squid,Loligo duvauceli Orbigny[J].J Food Sci Technol,2014,51(8):1509-16.
[31]COLOMBATTI A,BONALDO P,DOLIANA R.Type A modules:interacting domains found in several non-fibrillar collagens and in other extracellular matrix proteins[J].Matrix,1993,13(4):297-306.
[32]RUSKAMO S,YLNNE J.Structure of the human filamin A actin-binding domain[J].Acta Crystallogr D Biol Crystallogr,2009,65(11):1 217-1 221.
[33]HERRMANN H,B?R H,KREPLAK L,et al.Intermediate filaments:from cell architecture to nanomechanics[J].Nat Rev Mol Cell Biol,2007,8(7):562-573.
[34]EVANS J S.“Tuning in”to mollusk shell nacre-and prismatic-associated protein terminal sequence.Implications for biomineralization and the construction of high performance inorganic-organic composites[J].Chemical Reviews,2008,108:4 455-4462.