TY - JOUR
T1 - Evolution of Src Homology 2 (SH2) Domain to Recognize Sulfotyrosine
AU - Ju, Tong
AU - Niu, Wei
AU - Guo, Jiantao
N1 - Publisher Copyright:
© 2016 American Chemical Society.
PY - 2016/9/16
Y1 - 2016/9/16
N2 - Protein tyrosine O-sulfation is considered as the most common type of post-translational tyrosine modification in nature and plays important roles in extracellular biomolecular interactions. To facilitate the mapping, biological study, and medicinal application of this type of post-translational modification, we seek to evolve a small protein scaffold that recognizes sulfotyrosine with high affinity. We focused our efforts on the engineering of the Src Homology 2 (SH2) domain, which represents the largest class of known phosphotyrosine-recognition domain in nature and has a highly evolvable binding pocket. By using phage display, we successfully engineered the SH2 domain to recognize sulfotyrosine with high affinity. The best mutant, SH2-60.1, displayed more than 1700 fold higher sulfotyrosine-binding affinity than that of the wild-type SH2 domain. We also demonstrated that the evolved SH2 domain mutants could be used to detect sulfoprotein levels on the cell surface. These evolved SH2 domain mutants can be potentially applied to the study of protein tyrosine O-sulfation with proper experimental designs.
AB - Protein tyrosine O-sulfation is considered as the most common type of post-translational tyrosine modification in nature and plays important roles in extracellular biomolecular interactions. To facilitate the mapping, biological study, and medicinal application of this type of post-translational modification, we seek to evolve a small protein scaffold that recognizes sulfotyrosine with high affinity. We focused our efforts on the engineering of the Src Homology 2 (SH2) domain, which represents the largest class of known phosphotyrosine-recognition domain in nature and has a highly evolvable binding pocket. By using phage display, we successfully engineered the SH2 domain to recognize sulfotyrosine with high affinity. The best mutant, SH2-60.1, displayed more than 1700 fold higher sulfotyrosine-binding affinity than that of the wild-type SH2 domain. We also demonstrated that the evolved SH2 domain mutants could be used to detect sulfoprotein levels on the cell surface. These evolved SH2 domain mutants can be potentially applied to the study of protein tyrosine O-sulfation with proper experimental designs.
UR - http://www.scopus.com/inward/record.url?scp=84987804397&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84987804397&partnerID=8YFLogxK
U2 - 10.1021/acschembio.6b00555
DO - 10.1021/acschembio.6b00555
M3 - Article
C2 - 27428792
AN - SCOPUS:84987804397
SN - 1554-8929
VL - 11
SP - 2551
EP - 2557
JO - ACS chemical biology
JF - ACS chemical biology
IS - 9
ER -