2019/12/31 Incorporation of Unnatural Amino Acids - Profacgen

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Incorporation of Unnatural Amino Acids

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2019/12/31 Incorporation of Unnatural Amino Acids - Profacgen

Incorporation of Unnatural Amino Acids

The power of protein engineering has been largely expanded in recent years,
creating new possibilities in the use of unnatural amino acids (UAA) to interrogate or
construct reactivity, stability and spectral properties of engineered proteins.1
Experts at Profacgen provide the most up-to-date protein engineering technologies
for incorporation of UAAs bearing bioorthogonal handles such as:
Azides for click reaction or Staudinger ligation;
Alkynes for click reaction;
Ketones for amine-based reactions;
Aryl halides for Palladium-catalyzed cross-coupling;
Alkenes for metathesis.
Profacgen offers two strategies for UAA incorporation: 1) Metabolic incorporation of
azidohomoalanine or homopropargylglycine using native translational machinery;
2) Site-specific incorporation of UAAs by orthogonal suppressor tRNA/aminoacyl-
tRNA synthetase (aaRS) pairs.

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2019/12/31 Incorporation of Unnatural Amino Acids - Profacgen

Metabolic incorporation is a fast, sensitive, non-toxic and convenient way to

introduce methionine analogs (azidohomoalanine and homopropargylglycine) that
contain very small modifications during active protein synthesis. Azide- or alkyne-
modified proteins are labeling by exogenous reagents using either click reaction or
Staudinger ligation. Detection sensitivity in 1-D gels and Western blots is in the
femtomole range and suitable for downstream spectroscopic analysis (e.g., LC/MS).2

Site-specific incorporation utilizes tRNA/aaRS pair to deliver UAAs in response to a

nonsense or four-base (frameshift) codon. The engineered aaRS only recognize its
cognate tRNA without interfering endogenous tRNAs. On the other hand, the
orthogonal tRNA is not a substrate for native tRNA synthetases, but it can be
recognized by the engineered synthetase to transfer the specific UAA. This
approach has provided insights into questions that are otherwise impossible to
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2019/12/31 Incorporation of Unnatural Amino Acids - Profacgen

solve.
Profacgen, relying on the advanced protein engineering platform and over a decade
experience, offer a comprehensive set of UAA incorporation services to help our
clients:
Design customized research plans;
Synthesize UAAs with proper bioorthogonal handles;
Perform metabolic or site-specific incorporation of UAAS;
Purify and characterize modified proteins.
For more information regarding Profacgen’s UAA incorporation service, please
contact us (mailto:info@profacgen.com) or call 1-631-448-8149. Our customer
service representatives are available 24 hours a day, Monday through Friday, to
assist you.
References:
1. Lang, K.; et al. Cellular incorporation of unnatural amino acids and bioorthogonal
labeling of proteins. Chemical Reviews 2014, 114 (9): 4764-4806.
2. Beatty K.E.; et al. Noncanonical amino acids in protein science and engineering.
In: Köhrer C.; et al. (eds) Protein engineering. Nucleic acids and molecular biology
2009, (22), Springer, Berlin, Heidelberg

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