Engineering specific chemical modification sites into a collagen-like protein from Streptococcus pyogenes.
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.
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168, Australia.
Violet
V
Stoichevska
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168, Australia.
Yong Y
YY
Peng
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168, Australia.
Aditya V
AV
Vashi
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168, Australia.
Jerome A
JA
Werkmeister
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168, Australia.
Geoff J
GJ
Dumsday
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168, Australia.
John A M
JA
Ramshaw
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D003094:Collagen / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013297:Streptococcus pyogenes / Q000166:cytology / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D003094:Collagen
bacterial collagen; chemical modification; crosslinking; recombinant