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Protocol for resolving enzyme orientation and dynamics in advanced porous materials via SDSL-EPR.

ABSTRACT: Enzyme encapsulation in metal-organic frameworks (MOFs)/covalent-organic frameworks (COFs) provides advancement in biocatalysis, yet the structural basis underlying the catalytic performance is challenging to probe. Here, we present an effective protocol to determine the orientation and dynamics of enzymes in MOFs/COFs using site-directed spin labeling and electron paramagnetic resonance spectroscopy. The protocol is demonstrated using lysozyme and can be generalized to other enzymes. For complete information on the generation and use of this protocol, please refer to Pan et al. (2021a).

SUBMITTER: Pan Y

PROVIDER: S-EPMC8287244 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Protocol for resolving enzyme orientation and dynamics in advanced porous materials via SDSL-EPR.

Pan Yanxiong Y Li Hui H Li Qiaobin Q Lenertz Mary M Schuster Isabelle I Jordahl Drew D Zhu Xiao X Chen Bingcan B Yang Zhongyu Z

STAR protocols 20210714 3

Enzyme encapsulation in metal-organic frameworks (MOFs)/covalent-organic frameworks (COFs) provides advancement in biocatalysis, yet the structural basis underlying the catalytic performance is challenging to probe. Here, we present an effective protocol to determine the orientation and dynamics of enzymes in MOFs/COFs using site-directed spin labeling and electron paramagnetic resonance spectroscopy. The protocol is demonstrated using lysozyme and can be generalized to other enzymes. For comple ...[more]

PMID: 34308381

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