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Cellular synthesis of non-linear proteins

Zhang, Wen-Bin and Sun, Fei and Arnold, Frances H. and Tirrell, David A. (2013) Cellular synthesis of non-linear proteins. In: 246th ACS National Meeting & Exposition, Sept. 8-12, 2013, Indianapolis, IN.

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The spontaneous isopeptide bond formation between a peptide tag (SpyTag) and its protein partner (SpyCatcher) allows the synthesis of non-linear proteins in cellular hosts. Strategic placement of sequences encoding SpyTag and SpyCatcher within protein-coding genes programs the post-translational modification of the expressed proteins in situ, and enables the synthesis of a variety of unconventional protein topologies. For example, when SpyTag and SpyCatcher were placed at the Nand C-termini, resp., of an elastin-like-protein (ELP), highly efficient cyclization of the ELP was obsd. At low protein synthesis rates (e.g., at low temp. in relatively poor media), the product was almost exclusively monomeric circular ELP; at high protein synthesis rates (e.g., at high temp. in rich media), the major product was monomeric circular protein while significant amts. of chain-extended oligomers were also obsd. The circular topol. has been demonstrated by SDS-PAGE, MALDI-TOF mass spectrometry, proteolytic digestion, and single-site mutation.

Item Type:Conference or Workshop Item (Paper)
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Arnold, Frances H.0000-0002-4027-364X
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2013 American Chemical Society.
Record Number:CaltechAUTHORS:20131010-083215575
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:41849
Deposited By: Tony Diaz
Deposited On:10 Oct 2013 18:47
Last Modified:03 Oct 2019 05:52

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