Autonomous Synthesis of Fluorescent Silica Biodots Using Engineered Fusion Proteins

ACS Omega
Tolga T OlmezUrartu Ozgur Safak Seker

Abstract

Formation of biological materials is a well-controlled process that is orchestrated by biomolecules such as proteins. Proteins can control the nucleation and mineralization of biomaterials, thereby forming the hard tissues of biological organisms, such as bones, teeth, and shells. In this study, the design and implementation of multifunctional designer proteins are demonstrated for fluorescent silica micro/nanoparticle synthesis. The R5 motif of silaffin polypeptide, which is known for its silicification capability, was fused genetically into three spectrally distinct fluorescent proteins with the intention of forming modified fluorescent proteins. The bifunctional R5 peptide domain served as a tag to provide silica synthesis at ambient conditions. Three functional fusion constructs have been prepared, including GFPmut3-R5, Venus YFP-R5, and mCherry-R5. Recombinant fluorescent proteins were purified using silica-binding peptide tag through silica gel resin. Purified proteins were tested for their binding affinity to silica using quartz crystal microbalance with dissipation monitoring to make sure they can interact strong enough with the silica surfaces. Later, engineered fluorescent proteins were used to synthesize silica nano/...Continue Reading

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Citations

May 25, 2020·Acta Biomaterialia·Mohamed A A Abdelhamid, Seung Pil Pack
Mar 13, 2021·Biomaterials Science·Ebru Sahin KehribarUrartu Ozgur Safak Seker
Jan 2, 2021·Colloids and Surfaces. B, Biointerfaces·Esra YucaUrartu Özgür Şafak Şeker
Apr 15, 2021·Annual Review of Chemical and Biomolecular Engineering·Karthik PushpavanamFrançois Baneyx

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Datasets Mentioned

BETA
MDA-MB-436

Methods Mentioned

BETA
protein affinity purification
fluorescence microscopy
electrophoresis
Quartz crystal microbalance
fluorescence spectrometry
Fluorescence
electron microscopy
X-ray
biosensing
PCRs

Software Mentioned

ImageJ
IDT oligo analyzer
ExPASy

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