Analysis of protein interactions with two-hybrid system in cultured insect cells.
Many biological processes are usually coupled to the formation of protein complexes. The yeast two-hybrid system is a powerful tool for analyzing protein-protein interactions. Different patterns of protein modifications, such as glycosylation, phosphorylation, and acetylation, may affect the ability of proteins to interact. In this study, we developed the two-hybrid system that can be used in insect cells. To validate the insect two-hybrid (I2H) system, we analyzed and confirmed the known oligomer or dimer formation of silkworm Rad51 or RPA2-RPA3, respectively. The results established the feasibility of the I2H system for efficient analysis of protein interaction under conditions that closely reflect the normal physiological environment.
Mon
Hiroaki
H
Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka 812-8581, Japan.
Sugahara
Ryohei
R
Hong
Sun-Mee
SM
Lee
Jae-Man
JM
Kamachi
Yusuke
Y
Kawaguchi
Yutaka
Y
Kusakabe
Takahiro
T
D000818:Animals; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D019476:Insect Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D021122:Protein Subunits / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051135:Rad51 Recombinase / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D051716:Replication Protein A / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D019476:Insect Proteins; D021122:Protein Subunits; D051716:Replication Protein A; D051135:Rad51 Recombinase