Chimeric proteins

Figure 17.14 Model of evolved mutant from cephalosphorinase shuffling. The sequence of the most active cephalosporinase mutant was modeled using the crystal structure of the class C cephalosporinase from Enterobacter cloacae. The mutant and wild-type proteins were 63% identical. This chimeric protein contained portions from three of the starting genes, including Enterobacter (blue), Klebsiella (yellow), and Citrobacter (green), as well as 33 point mutations (red). (Courtesy of A. Crameri.)...

Maeda, Y., et al. (1997). Engineering of functional chimeric protein G-Vargula luciferase. Anal. Biochem. 249 147-152. 

Keywords Amphiphilic polymer Cell adhesion Chimeric protein Islet of Langerhans Self-assembled monolayer Stem cell... 

Fig. 6 EGF-containing chimeric proteins anchored to the Ni-chelated surface through coordination. Bold lines in the molecular structures represent chelate bonding. TEG-thiol triethylene glycol-containing alkanethiol. Reproduced from Nakaji-Hirabayashi et al. [87] with permission from American Chemical Society, copyright 2009...

Numerous studies have shown that EGF binding to EGFR triggers receptor dimerization. This is considered a crucial step in intracellular signal transduction [98]. Inspired by this mechanism, we designed EGF chimeric proteins that spontaneously dimerized (dEGF-His). These dimers were terminally anchored to the substrate. We expected that these preformed dimeric EGF structures would facilitate the formation of EGF-EGFR dimer complexes more efficiently than monomeric EGF structures. 

Our strategy for the spontaneous dimerization of EGF was to incorporate an ot-helical oligopeptide with the ability to participate in forming a coiled-coil structure [99]. We implemented a heterodimerization system in which (KELASVK)5 (K5) and (EKLASVE)5 (E5) peptides could form stable coiled-coil heterodimers. We then synthesized two chimeric proteins that contained EGF attached to either the K5 (EGF-K5-His) or the E5 (EGF-E5-His). Both had the hexahistidine sequence added to the C-terminus for anchoring through coordination with Ni2+ ions fixed to a substrate. 

Davio, S.R., K.M. Kienle, and B.E. Collins. 1995. Interdomain interactions in the chimeric protein toxin sCD4(178)-PE40 a differential scanning calorimetry (DSC) study. Pharm Res 12 642-648. 

An intensely fluorescent protein isolated from the jellyfish Aequorea victoria and other Aequorea species. This protein produces its fluorescence from an intrinsic chro-mophore generated by a series of steps involving residues 65-67 of the polypeptide chain. As a result, GFP has broad application, especially when expressed as a fused or chimeric protein containing a second polypeptide region whose intracellular location is of interest. ... 

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