Structural characterization of amorphous calcium carbonate-binding protein: an insight into the mechanism of amorphous calcium carbonate formation

被引:24
作者
Su, Jingtan [1 ]
Liang, Xiao [1 ]
Zhou, Qiang [2 ]
Zhang, Guiyou [1 ]
Wang, Hongzhong [1 ]
Xie, Liping [1 ,3 ]
Zhang, Rongqing [1 ,3 ]
机构
[1] Tsinghua Univ, Sch Life Sci, Inst Marine Biotechnol, Beijing 100084, Peoples R China
[2] Tsinghua Univ, Sch Life Sci, State Key Lab Biomembrane & Membrane Biotechnol, Beijing 100084, Peoples R China
[3] Tsinghua Univ, Minist Educ, Prot Sci Lab, Beijing 10084, Peoples R China
基金
中国国家自然科学基金; 国家高技术研究发展计划(863计划);
关键词
amorphous calcium carbonate (ACC); biomineralization; homology modelling; oligomerization; structure-function relationship; symmetry; PRECURSOR PHASE; SHELL FORMATION; MATRIX PROTEIN; PEARL OYSTER; CRYSTALLIZATION; NACRE; MODEL; NUCLEATION; OVALBUMIN; BIOMINERALIZATION;
D O I
10.1042/BJ20130285
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
ACC (amorphous calcium carbonate) plays an important role in biomineralization process for its function as a precursor for calcium carbonate biominerals. However, it is unclear how biomacromolecules regulate the formation of ACC precursor in vivo. In the present study, we used biochemical experiments coupled with bioinformatics approaches to explore the mechanisms of ACC formation controlled by ACCBP (ACC-binding protein). Size-exclusion chromatography, chemical cross-linking experiments and negative staining electron microscopy reveal that ACCBP is a decamer composed of two adjacent pentamers. Sequence analyses and fluorescence quenching results indicate that ACCBP contains two Ca2+-binding sites. The results of in vitro crystallization experiments suggest that one Ca2+-binding site is critical for ACC formation and the other site affects the ACC induction efficiency. Homology modelling demonstrates that the Ca2+-binding sites of pentameric ACCBP are arranged in a 5-fold symmetry, which is the structural basis for ACC formation. To the best of our knowledge, this is the first report on the structural basis for protein-induced ACC formation and it will significantly improve our understanding of the amorphous precursor pathway.
引用
收藏
页码:179 / 186
页数:8
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