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Engineering specific chemical modification sites into a collagen‐like protein from Streptococcus pyogenes
Author(s) -
Stoichevska Violet,
Peng Yong Y.,
Vashi Aditya V.,
Werkmeister Jerome A.,
Dumsday Geoff J.,
Ramshaw John A. M.
Publication year - 2017
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.35957
Subject(s) - recombinant dna , chemical modification , azide , cysteine , reagent , bifunctional , combinatorial chemistry , streptococcus pyogenes , escherichia coli , click chemistry , biochemistry , materials science , residue (chemistry) , yield (engineering) , amino acid , bacteria , chemistry , organic chemistry , biology , enzyme , genetics , metallurgy , gene , staphylococcus aureus , catalysis
Recombinant bacterial collagens provide a new opportunity for safe biomedical materials. They are readily expressed in Escherichia coli in good yield and can be readily purified by simple approaches. However, recombinant proteins are limited in that direct secondary modification during expression is generally not easily achieved. Thus, inclusion of unusual amino acids, cyclic peptides, sugars, lipids, and other complex functions generally needs to be achieved chemically after synthesis and extraction. In the present study, we have illustrated that bacterial collagens that have had their sequences modified to include cysteine residue(s), which are not normally present in bacterial collagen‐like sequences, enable a range of specific chemical modification reactions to be produced. Various model reactions were shown to be effective for modifying the collagens. The ability to include alkyne (or azide) functions allows the extensive range of substitutions that are available via “click” chemistry to be accessed. When bifunctional reagents were used, some crosslinking occurred to give higher molecular weight polymeric proteins, but gels were not formed. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 806–813, 2017.