Enamel is the covering tissue of teeth, made of regularly arranged hydroxyapatite crystals deposited on an organic matrix composed of 90% amelogenin that is completely degraded at the end of the enamel formation process. Amelogenin has a biomineralizing activity, forming nanoparticles or nanoribbons that guide hydroxyapatite deposit, and regenerative functions in bone and vascular tissue and in wound healing. Biotechnological products containing amelogenin seem to facilitate these processes. Here, we describe the production of human amelogenin in plants by transient transformation of Nicotiana benthamiana with constructs carrying synthetic genes with optimized human or plant codons. Both genes yielded approximately 500 µg of total amelogenin per gram of fresh leaf tissue. Two purification procedures based on affinity chromatography or on intrinsic solubility properties of the protein were followed, yielding from 12 to 150 µg of amelogenin per gram of fresh leaf tissue, respectively, at different purity. The identity of the plant-made human amelogenin was confirmed by MALDI-TOF-MS analysis of peptides generated following chymotrypsin digestion. Using dynamic light scattering, we showed that plant extracts made in acetic acid containing human amelogenin have a bimodal distribution of agglomerates, with hydrodynamic diameters of 22.8 ± 3.8 and 389.5 ± 86.6 nm. To the best of our knowledge, this is the first report of expression of human amelogenin in plants, offering the possibility to use this plant-made protein for nanotechnological applications.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/s12033-017-0030-y | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Disclosure of biological sex may impact individual privacy.
Forensic Sci Int Genet
December 2024
Department of Forensic Medicine, University of Helsinki, Helsinki, Finland; Forensic Science Institute, Radford University, Radford, VA, USA. Electronic address:
Human identification by forensic DNA profiling primarily relies on the analysis of short tandem repeat markers (STRs) and Amelogenin or other sex determining markers. The resultant DNA profiles can be compared directly between evidence and reference samples or indirectly (i.e.
View Article and Find Full Text PDFPeroxisomal dysfunction interferes with odontogenesis and leads to developmentally delayed teeth and defects in distinct dental cells in Pex11b-deficient mice.
PLoS One
December 2024
Institute of Anatomy and Cell Biology, Justus-Liebig-University, Giessen, Germany.
Human peroxisomal biogenesis disorders of the Zellweger syndrome spectrum affect skeletal development and induce tooth malformations. Whereas several peroxisomal knockout mouse studies elucidated the pathogenesis of skeletal defects, little information is available on how dental pathologies arise in peroxisomal biogenesis disorder patients. To understand the impact of severe peroxisomal dysfunction on early odontogenesis, here we performed morphometric studies on developing molars of new-born Pex11b knockout mice.
View Article and Find Full Text PDFLAP-MALDI MS analysis of amelogenin from teeth for biological sex estimation.
J Pharm Biomed Anal
November 2024
Department of Chemistry, University of Reading, Whiteknights, Reading RG6 6DX, United Kingdom. Electronic address:
The biological sex estimation of human individuals can be achieved by extracting fragments of the amelogenin protein from small areas of tooth enamel. The amelogenin gene can be found on both sex chromosomes (X and Y) with chromosome-specific differences in its sequence, and consequently the sequences of the expressed protein in teeth. Virtually all current analytical techniques used to identify the occurrence of the male Y chromosome-specific proteoform employ proteoform-specific peptide analysis by LC-ESI MS/MS, which typically results in longer analytical times due to the LC separation step, despite recent efforts of shortening the LC step.
View Article and Find Full Text PDFInitial Clinical and Cone-Beam Computed Tomography (CBCT) Outcomes of Enamel Matrix Derivative-Based Regenerative Therapy in Vertical Bone Defects.
Cureus
October 2024
Department of Periodontology and Dental Implantology, Medical University of Varna, Varna, BGR.
Background and objective Enamel matrix derivatives (EMD) are biomaterials extracted from porcine dental germs. They consist of amelogenins (90%), ameloblastine, enamelin, and amelotin. New evidence has shown that all these proteins can be absorbed onto hydroxyapatite crystals and collagen fibers on the root surface of teeth and initiate regeneration of root cementum as well as stimulate periodontal cell proliferation.
View Article and Find Full Text PDFConcurrent genotyping of mitochondrial DNA and nuclear DNA in rootless hair shafts and blood samples for enhanced analysis.
Forensic Sci Int Genet
February 2025
Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China; Guangdong Province Translational Forensic Medicine Engineering Technology Research Center, Sun Yat-sen University, Guangzhou 510080, China. Electronic address:
Article Synopsis
- - Hair is a crucial source of biological evidence in crime scenes, but extracting and analyzing DNA from hair shafts can be difficult due to the degradation of DNA, which makes traditional methods less effective.
- - A new all-in-one multiplex system, the MGIEasy Signature Identification Library Prep Kit, allows for the simultaneous analysis of mitochondrial DNA (mtDNA) and nuclear DNA (nuDNA) using advanced sequencing technology, showing promising results from a study involving 370 hair shafts and 180 blood samples.
- - Findings revealed significant genetic variations in the population studied, with differences in DNA extraction methods impacting the efficiency of nuDNA genotyping, and a notable decrease in the depth of cover for mtDNA as the hair shaft length increased. *
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!