High Expression of Human Amelogenin in E. Coli

Deutsch, D.; Chityat, E.; Hekmati, M; Palmon, Aaron; Farkash, Yigal; Dafni, Leah

doi:10.1177/08959374960100021201

Cited by 7 publications

(4 citation statements)

References 48 publications

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“…Expression of rhAm has been reported in different expression systems , whereas, there has been no previous report so far, on phosphorylation state of these recombinant proteins. Taylor et al .…”

Section: Discussionmentioning

confidence: 99%

Analogous effects of recombinant human full‐length amelogenin expressed by Pichia pastoris yeast and enamel matrix derivative in vitro

et al. 2012

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Section: Discussionmentioning

confidence: 99%

Analogous effects of recombinant human full‐length amelogenin expressed by Pichia pastoris yeast and enamel matrix derivative in vitro

et al. 2012

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“…Amelogenin is a hydrophobic enamel matrix protein that possesses the unique physiological characteristic of self-aggregation into nano -spheres, an important structural feature related to its function in the developing enamel; but one that makes it challenging to express, isolate, and analyze [3]. Expression of a recombinant amelogenin protein has been reported in prokaryotic systems mostly [18][19][20][21]. These recombinant amelogenin proteins have been used in various structural and functional studies [23].…”

Section: Discussionmentioning

confidence: 99%

“…This problem has been previously overcome by high expression of mouse amelogenin [18], porcine amelogenin [19] and human amelogenin [20] in prokaryotic bacterial systems. In this research, we describe the expression of a recombinant human amelogenin protein in the prokaryotic bacterial systems, which has made possible the investigation of novel functions of amelogenin by providing relatively high amounts of pure amelogenin, free from contamination of other enamel matrix proteins.…”

Section: Introductionmentioning

confidence: 99%

“…During enamel development and mineralization, the secreted amelogenins are lost from the tissue,along with most of the other enamel matrix proteins, due to sequential degradation by specific proteases such as enamelysin (matrix metalloproteinase 20, and kallikrein 4 (Klk4) [6]. They are replaced by mineral ions, calcium, and phosphorus, which eventually results in fully mineralized, hard, and mature enamel [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

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Expression and Purification of Recombinant Human Amelogenin Using the Prokaryotic Expression System

Feng¹,

Yao²,

Du³

et al. 2016

Proceedings of the 2016 International Conference on Advanced Materials Science and Environmental Engineering

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Abstract-The amelogenins (Am) constitute about 90% of the enamel matrix proteins and play a critical role in enamel biomineralization. This study aimed to express and characterize recombinant human amelogenin (rAm) protein in the prokaryotic expression system in quantities sufficient for structural and functional research. Human cDNA coding for a 175 amino acid amelogenin protein was subcloned into the pET-28a(+) vector, this system adds a hexa-histidine tag to N-terminal amino acid of the expressed protein, enabling effective three steps purification by Ni

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Cloning and Expression Analysis of Human Amelogenin in Nicotiana benthamiana Plants by Means of a Transient Expression System

et al. 2017

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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.

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High Expression of Human Amelogenin in E. Coli

Cited by 7 publications

References 48 publications

Analogous effects of recombinant human full‐length amelogenin expressed by Pichia pastoris yeast and enamel matrix derivative in vitro

Analogous effects of recombinant human full‐length amelogenin expressed by Pichia pastoris yeast and enamel matrix derivative in vitro

Expression and Purification of Recombinant Human Amelogenin Using the Prokaryotic Expression System

Cloning and Expression Analysis of Human Amelogenin in Nicotiana benthamiana Plants by Means of a Transient Expression System

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