Study of human leucine-rich amelogenin peptide and its regulation of mineralization by cryogenic transmission electron microscopy.
- VernacularTitle:人釉原蛋白亮氨酸富集片段的冷冻电子显微镜观察及其引导矿化性能研究
- Author:
Kun TIAN
1
;
Xiaoyun FENG
2
Author Information
- Publication Type:Journal Article
- Keywords: amelogenin; cryogenic transmission electron microscopy; hydroxyapatite; leucine-rich amelogenin peptide
- MeSH: Amelogenin; Bone Density; Calcium Phosphates; Crystallization; Dental Enamel; Dental Enamel Proteins; Durapatite; Humans; Microscopy, Electron, Transmission
- From: West China Journal of Stomatology 2017;35(1):63-67
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVERecombinant human leucine-rich amelogenin peptide (LRAP) was studied by cryogenic transmission electron microscopy (TEM); evaluation focused on its self-assembly and crystal growth in vitro.
METHODSHuman LRAP was recombined through prokaryotic expression vector pCold-SUMO and transformed into Escherichia coli BL21plys to acquire purified proteins. Cryogen TEM recorded assembly and self-assembling of LRAP from pH 3.5 to pH 8.0, and the hydroxyapatite crystal growth in the mixture of LRAP protein solution and artificial saliva was observed using TEM and selected area electron diffraction.
RESULTSMore than 90% purity LRAP was expressed, purified and identified as described in methods. LRAP linked into oligomers, nanospheres, nanochains, and microribbons, whereas pH value increased from 3.5 to 8.0. Mature hydroxyapatite crystal growth was guided in artificial saliva filled with calcium phosphate.
CONCLUSIONSLRAP is simplified amelogenin functional domain and conserved the basic characters of amelogenin such as self-assembling and inducing crystallization along c axis. In the area of acellular synthesis of hydroxyapatite using extracellular enamel matrix protein, LRAP is one of candidate repair materials for irregular hard tissue defection. .