title: Display of membrane proteins on the heterologous caveolae carved by caveolin-1 in the Escherichia coli cytoplasm.
abstract: Caveolae are membrane-budding structures that exist in many vertebrate cells. One of the important functions of caveolae is to form membrane curvature and endocytic vesicles. Recently, it was shown that caveolae-like structures were formed in Escherichia coli through the expression of caveolin-1. This interesting structure seems to be versatile for a variety of biotechnological applications. Targeting of heterologous proteins in the caveolae-like structure should be the first question to be addressed for this purpose. Here we show that membrane proteins co-expressed with caveolin-1 are embedded into the heterologous caveolae (h-caveolae), the cavaolae-like structures formed inside the cell. Two transmembrane SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins, Syntaxin 1a and vesicle-associated membrane protein 2 (VAMP2), were displayed on the h-caveolae surface. The size of the h-caveolae harboring the transmembrane proteins was ∼100 nm in diameter. The proteins were functional and faced outward on the h-caveolae. Multi-spanning transmembrane proteins FtsH and FeoB could be included in the h-caveolae, too. Furthermore, the recombinant E. coli cells were shown to endocytose substrate supplemented in the medium. These results provide a basis for exploiting the h-caveolae formed inside E. coli cells for future biotechnological applications.
authors: Department of Genetic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 440-746, South Korea; Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon, Gyeonggi-do 440-746, South Korea.
authors: Jonghyeok
authors: J
authors: Shin
authors: Department of Genetic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 440-746, South Korea; Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon, Gyeonggi-do 440-746, South Korea.
authors: Young-Hun
authors: YH
authors: Jung
authors: Department of Genetic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 440-746, South Korea; Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon, Gyeonggi-do 440-746, South Korea.
authors: Da-Hyeong
authors: DH
authors: Cho
authors: Department of Genetic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 440-746, South Korea; Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon, Gyeonggi-do 440-746, South Korea.
authors: Myungseo
authors: M
authors: Park
authors: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, South Korea.
authors: Kyung Eun
authors: KE
authors: Lee
authors: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, South Korea.
authors: Yoosoo
authors: Y
authors: Yang
authors: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, South Korea.
authors: Cherlhyun
authors: C
authors: Jeong
authors: Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, South Korea.
authors: Bong Hyun
authors: BH
authors: Sung
authors: Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, South Korea.
authors: Jung-Hoon
authors: JH
authors: Sohn
authors: Department of Food Science and Engineering, Ewha Womans University, Seoul 120-750, South Korea.
authors: Jin-Byung
authors: JB
authors: Park
authors: Department of Genetic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 440-746, South Korea; Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon, Gyeonggi-do 440-746, South Korea. Electronic address: dhkweon@skku.edu.
authors: Dae-Hyuk
authors: DH
authors: Kweon
mesh_terms: D049069:ATP-Dependent Proteases / Q000378:metabolism; D001709:Biotechnology; D027682:Cation Transport Proteins / Q000378:metabolism; D021941:Caveolae / Q000378:metabolism / Q000648:ultrastructure; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004705:Endocytosis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D050827:Syntaxin 1 / Q000235:genetics / Q000378:metabolism; D050988:Vesicle-Associated Membrane Protein 2 / Q000235:genetics / Q000378:metabolism
chemical_list: C495540:CAV1 protein, human; D027682:Cation Transport Proteins; D051242:Caveolin 1; D029968:Escherichia coli Proteins; C470580:FeoB protein, E coli; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D050827:Syntaxin 1; D050988:Vesicle-Associated Membrane Protein 2; D049069:ATP-Dependent Proteases; C513383:FtsH protein, E coli
keywords: Caveolin-1; Heterologous caveolae; Transmembrane protein