DENTIN BIOMINERALIZATION

牙本质生物矿化

• 批准号: 7231485
• 负责人: CHARLES SFEIR
• 金额: $ 28.31万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7231485
• 关键词: Address; Adopted; Affect; Apatites; Applications Grants; Atomic Force Microscopy; Biological; C-terminal; Calcium Binding; Cell Line; Cells; Characteristics; Charge; Chemicals; Chimeric Proteins; Classification; Code; Collagen; Collagen Type I; Condition; Core Facility; Crystal Formation; Data; Dentin; Dentin Formation; Deposition; Disease; Electron Microscopy; Electron energy loss spectroscopy; Environment; Escherichia; Event; Extracellular Matrix Proteins; Fibroblasts; Future; Genes; Goals; Growth; Habits; Heart Valve Prosthesis; In Vitro; Invertebrates; Kidney Calculi; Kinetics; Knowledge; Learning; Localized; Mammalian Cell; Mediating; Minerals; Modeling; Modification; Nature; Odontoblasts; Osteoblasts; Pattern; Phosphoproteins; Phosphorylation; Phosphorylation Site; Physiologic Ossification; Physiological; Play; Post-Translational Protein Processing; Process; Property; Protein Sequence Analysis; Proteins; Proteomics; Reaction; Recombinants; Regulation; Relative (related person); Research Proposals; Resolution; Rest; Role; Scanning; Scanning Transmission Electron Microscopy Procedures; Serine; Shapes; Site; Specific qualifier value; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis; Structure; Surface; System; Testing; Time; Tissues; Universities; Vertebrates; Work; biomineralization; bone; calcification; casein kinase II; concept; dentinal phosphophoryn; extracellular; in vitro Model; in vivo; in vivo Model; inorganic phosphate; interfacial; macromolecule; mass spectrometer; mineralization; mutant; novel; phosphophoryn; research study; secretory protein; size; therapy design

DESCRIPTION (provided by applicant): Biomineralization is one of the most widespread and important processes in both vertebrates and invertebrates. We have directed our efforts to study the mineralization of bone and dentin in which apatite crystals are specifically grown within type I collagen matrices. Although it is likely that the overall mechanisms involved in mineralization of the two tissues are similar, we have adopted the dentin system since it represents a relatively simpler system than bone. The basic working hypothesis has been that certain acidic, phosphorylated, non-collagenous extracellular matrix protein macromolecules (NCP) are first localized by interaction with the collagen I fibril surfaces, which then direct the nucleation of crystal deposits and orientation of certain specific crystallographic habit directions relative to the fibril axes. Further interactions of the NCP with the growing crystals are postulated to specifically regulate crystal growth in terms of habit, shape and size. Although there are many NCP present in dentin, we have focused our efforts on the primary phosphoprotein of dentin: Phosphophoryn ([PP]). PP is very acidic in nature and characterized by a very high content of serine residues, at about an 85 percent level of phosphorylation. In this proposal we plan to introduce the gene encoding for PP to over-express it in fibroblast, osteoblast and odontoblast cells, respectively. This will enable us to establish an in vivo system to study biomineralization. We will characterize the mineralization of the crystals in these three separate cells to ascertain the nucleation and crystal growth directions. This approach will allow us to address many important questions: Is the post-translationnal modification of PP similar in fibroblast in contrast to osteoblast and/or odontoblast cells? Will the mineralized crystal have the same crystallographic properties when the nucleating protein, such as PP, is expressed in a fibroblast without the rest of the NCP, which is usually expressed in an osteoblast or odontoblast? We will also study the effect of phosphorylation on the Mineralization process in vitro by expressing the recombinant PP and studying its in vitro phosphorylation on the mineral. These studies should yield new information related to the specific role of phosphoproteins in biomineralization. This knowledge will be extremely valuable in determining the mechanisms involved in the biomineralization process particularly since the studies are conducted in an in vivo model in comparison to the traditional approaches of in vitro studies of biomineralization. This data will also provide basic knowledge for designing treatment of ectopic mineralization specifically in diseases as such kidney stones or failing heart valve prosthesis.

描述（由申请人提供）：生物矿化是脊椎动物和无脊椎动物中最广泛和最重要的过程之一。我们指导了研究骨和牙本质矿化的努力，其中磷灰石晶体在I型胶原蛋白矩阵中特异性生长。尽管两种组织矿化涉及的总体机制可能相似，但我们采用了牙本质系统，因为它代表了一个相对简单的系统。基本的工作假设是，某些酸性，磷酸化的，非胶原性细胞外基质蛋白大分子（NCP）首先是通过与胶原蛋白I原纤维表面相互作用来定位的，然后将其指导晶体沉积物的成核和相对特定晶体学疗法型纤维轴的方向。假定NCP与生长晶体的进一步相互作用，以根据习惯，形状和大小特异性调节晶体生长。尽管牙本质中存在许多NCP，但我们将精力集中在牙本质的原发性磷酸蛋白上：磷phophophoryn（[PP]）。 PP本质上非常酸性，其特征是丝氨酸残基的含量很高，约为85％的磷酸化水平。 在此提案中，我们计划在成纤维细胞，成骨细胞和odontoblast细胞中引入PP编码的基因编码。这将使我们能够建立一个研究生物矿化的体内系统。我们将表征这三个独立细胞中晶体的矿化，以确定成核和晶体生长方向。这种方法将使我们能够解决许多重要的问题：与成骨细胞和/或odontoblast细胞相比，成纤维细胞中PP的翻译后修饰是否相似？当成核蛋白（例如PP）在没有其余NCP的成纤维细胞中表达成核蛋白（例如PP）时，矿化晶体是否具有相同的晶体学特性，而NCP的其余部分通常在成骨细胞或odontoblast中表达？我们还将通过表达重组PP并研究其在矿物质上的体外磷酸化来研究体外磷酸化对矿化过程的影响。 这些研究应产生与磷蛋白在生物矿化中的特定作用有关的新信息。这些知识对于确定生物矿化过程中涉及的机制将非常有价值，特别是因为与传统的生物矿化研究相比，研究是在体内模型中进行的。这些数据还将提供基本知识，用于设计异位矿化的治疗，专门针对此类肾结石或心脏瓣膜假体的疾病。

项目成果

Primary structure and phosphorylation of dentin matrix protein 1 (DMP1) and dentin phosphophoryn (DPP) uniquely determine their role in biomineralization.

• DOI: 10.1021/bm2005214
• 发表时间: 2011-08-08
• 期刊: BIOMACROMOLECULES
• 影响因子: 6.2
• 作者: Deshpande, Atul Suresh;Fang, Ping-An;Zhang, Xiaoyuan;Jayaraman, Thottala;Sfeir, Charles;Beniash, Elia
• 通讯作者: Beniash, Elia

Possible role of DMP1 in dentin mineralization.

• DOI: 10.1016/j.jsb.2010.11.013
• 发表时间: 2011-04
• 期刊: JOURNAL OF STRUCTURAL BIOLOGY
• 影响因子: 3
• 作者: Beniash, Elia;Deshpande, Atul S.;Fang, Ping An;Lieb, Nicholas S.;Zhang, Xiaoyuan;Sfeir, Charles S.
• 通讯作者: Sfeir, Charles S.

Cell derived hierarchical assembly of a novel phosphophoryn-based biomaterial.

新型基于磷酸基的生物材料的细胞衍生分层组装。

• DOI: 10.1159/000158571
• 发表时间: 2009
• 期刊: Cells, tissues, organs
• 作者: Li,Jinhua;Olton,Dana;Lee,Donghyun;Kumta,PrashantN;Sfeir,Charles
• 通讯作者: Sfeir,Charles

Synthesis of bone-like nanocomposites using multiphosphorylated peptides.

• DOI: 10.1016/j.actbio.2014.01.007
• 发表时间: 2014-05
• 期刊: ACTA BIOMATERIALIA
• 影响因子: 9.7
• 作者: Sfeir, Charles;Fang, Ping-An;Jayaraman, Thottala;Raman, Aparna;Zhang Xiaoyuan;Beniash, Elia
• 通讯作者: Beniash, Elia