Bioinspired Artificial Protein Materials: Self-Assembly and Order from Nano to Macroscale
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Bioinspired Artificial Protein Materials: Self-Assembly and Order from Nano to Macroscale Min Dai,1 Jennifer S. Haghpanah,1 Carlo Yuvienco,1 and Jin Kim Montclare1,2 Polytechnic Institute of New York University, 6 Metrotech Center, Brooklyn, NY, 11201 U.S.A. 2 SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, U.S.A. 1
ABSTRACT We describe the biosynthesis and characterization of protein materials comprised of two distinct self-assembling domains (SADs): elastin (E) found in tissue for its elastic properties and cartilage oligomeric matrix protein coiled-coil (COMPcc, C) predominantly located in joint and in bones. Based on earlier studies on protein block polymers comprised these two SADs, orientation and number of blocks play a crucial role in the overall stimuli-responsive supramolecular assembly behavior. Here we fabricate a range of EnC and CEn block polymers in which the E domain is systematically truncated to explore the effects of the E domain on the overall physicochemical behavior. INTRODUCTION The fabrication of multifunctional nano-materials that can self-assemble into defined structures bear tremendous potential in nanoelectronics, microarrays, drug delivery and regenerative medicine [1-9]. Recent advances in molecular and synthetic biology have brought protein polymers to the forefront as useful biomaterials with a broad range of physicochemical properties [10, 11]. Proteins not only provide a diversity of chemical functionality as the building blocks are comprised of 20 amino acids, but also present important 3-dimensional structures that provide order on the nano- to macro- length scales. Here, we present the fabrication and characterization of protein block polymers comprised of two distinct self-assembling domains (SADs) derived from elastin (E) and cartilage oligomeric matrix protein coiled-coil (C). Recently, three protein block polymers EC, CE and ECE have been biosynthesized and characterized. The orientation and number of blocks affect the overall secondary structure, stability and supramolecular assembly. Based on these studies, we choose to focus and further elaborate on the two diblocks EC and CE. To modulate the overall physicochemical properties of the protein block polymers, we perform a systematic truncation of the E domain for both protein diblocks yielding a library of EnC and CEn where n represents the number of E repeats (Figure 1).
Figure 1. Illustration of the EnC and CEn protein block polymers.
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EXPERIMENT Gene Synthesis. The following primers were used to amplify the library of E domain length from pUC19ELP pentamer (gift from D. Tirrell) plasmid: BamHI:5’ggaggccGGATCCaagccgattgcggctagcgcggtgccgg-3’, SacI:5’gccccGAGCTCcgatccctcgagcggcaccccgac-3’, SalI:5’ggaggccGTCGACaagccgattgcggctagcgcggtgccgg-3’, and HindIIII: 5’ggccccAAGCTTcgcaccggtacccgatccctcgagcggcaccccgac-3’. Fragments of E from 150 to 400 bps were amplified, gel purified and restricted with appropriate restriction enzymes. E inserts were cloned into the pQE30/C between restriction sites BamHI
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