Direct protein-protein conjugation by genetically introducing bioorthogonal functional groups into proteins.
Proteins often function as complex structures in conjunction with other proteins. Because these complex structures are essential for sophisticated functions, developing protein-protein conjugates has gained research interest. In this study, site-specific protein-protein conjugation was performed by genetically incorporating an azide-containing amino acid into one protein and a bicyclononyne (BCN)-containing amino acid into the other. Three to four sites in each of the proteins were tested for conjugation efficiency, and three combinations showed excellent conjugation efficiency. The genetic incorporation of unnatural amino acids (UAAs) is technically simple and produces the mutant protein in high yield. In addition, the conjugation reaction can be conducted by simple mixing, and does not require additional reagents or linker molecules. Therefore, this method may prove very useful for generating protein-protein conjugates and protein complexes of biochemical significance.
Department of Chemistry, Sogang University, Seoul 121-742, Republic of Korea.
Sanggil
S
Kim
Department of Chemistry, Sogang University, Seoul 121-742, Republic of Korea.
Wooseok
W
Ko
Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
Bong Hyun
BH
Sung
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Republic of Korea.
Sun Chang
SC
Kim
Department of Chemistry, Sogang University, Seoul 121-742, Republic of Korea. Electronic address: hslee76@sogang.ac.kr.
Hyun Soo
HS
Lee
D000596:Amino Acids / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D015394:Molecular Structure; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000596:Amino Acids; D011506:Proteins
Click reaction; Genetic incorporation; Protein–protein conjugation; Unnatural amino acids