TY - JOUR
T1 - Structural Characterization of Native and Modified Encapsulins as Nanoplatforms for in Vitro Catalysis and Cellular Uptake
AU - Putri, Rindia M.
AU - Allende-Ballestero, Carolina
AU - Luque, Daniel
AU - Klem, Robin
AU - Rousou, Katerina Asteria
AU - Liu, Aijie
AU - Traulsen, Christoph H.H.
AU - Rurup, W. Frederik
AU - Koay, Melissa S.T.
AU - Castón, José R.
AU - Cornelissen, Jeroen J.L.M.
N1 - Open Access
PY - 2017/12/26
Y1 - 2017/12/26
N2 - Recent years have witnessed the emergence of bacterial semiorganelle encapsulins as promising platforms for bio-nanotechnology. To advance the development of encapsulins as nanoplatforms, a functional and structural basis of these assemblies is required. Encapsulin from Brevibacterium linens is known to be a protein-based vessel for an enzyme cargo in its cavity, which could be replaced with a foreign cargo, resulting in a modified encapsulin. Here, we characterize the native structure of B. linens encapsulins with both native and foreign cargo using cryo-electron microscopy (cryo-EM). Furthermore, by harnessing the confined enzyme (i.e., a peroxidase), we demonstrate the functionality of the encapsulin for an in vitro surface-immobilized catalysis in a cascade pathway with an additional enzyme, glucose oxidase. We also demonstrate the in vivo functionality of the encapsulin for cellular uptake using mammalian macrophages. Unraveling both the structure and functionality of the encapsulins allows transforming biological nanocompartments into functional systems.
AB - Recent years have witnessed the emergence of bacterial semiorganelle encapsulins as promising platforms for bio-nanotechnology. To advance the development of encapsulins as nanoplatforms, a functional and structural basis of these assemblies is required. Encapsulin from Brevibacterium linens is known to be a protein-based vessel for an enzyme cargo in its cavity, which could be replaced with a foreign cargo, resulting in a modified encapsulin. Here, we characterize the native structure of B. linens encapsulins with both native and foreign cargo using cryo-electron microscopy (cryo-EM). Furthermore, by harnessing the confined enzyme (i.e., a peroxidase), we demonstrate the functionality of the encapsulin for an in vitro surface-immobilized catalysis in a cascade pathway with an additional enzyme, glucose oxidase. We also demonstrate the in vivo functionality of the encapsulin for cellular uptake using mammalian macrophages. Unraveling both the structure and functionality of the encapsulins allows transforming biological nanocompartments into functional systems.
KW - bacterial compartments
KW - cryo-electron microscopy
KW - encapsulin
KW - nanoplatforms
KW - nanoreactors
UR - http://www.scopus.com/inward/record.url?scp=85040078468&partnerID=8YFLogxK
U2 - 10.1021/acsnano.7b07669
DO - 10.1021/acsnano.7b07669
M3 - Article
AN - SCOPUS:85040078468
VL - 11
SP - 12796
EP - 12804
JO - ACS nano
JF - ACS nano
SN - 1936-0851
IS - 12
ER -