Skip to main content

Cryo-EM structure of the MMOH-MMOR complex


EMSL Project ID
51187

Abstract

Methane to methanol conversion is critical for environmental health, reducing greenhouse gas, and utilizing the natural gas as an alternative energy source. Methanotrophic bacteria utilize the methane (CH4) as a sole source of carbon and energy by oxidizing and breaking the highly stable C-H bond. This unique reaction can be achieved via the enzyme complex called methane monooxygenase (MMO).
Soluble methane monooxygenase (sMMO) is a multi-component protein complex comprising a 245-kDa (???)2 hydroxylase MMOH, a 16-kDa regulatory protein MMOB, a 38-kDa electron-transfer reductase MMOR, and 12-kDa potential MMO inhibitory subunit (MMOD). Since the maximum catalytic activity of sMMO is only achieved via the interplay of MMOH with three auxiliary subunits, structural elucidation is a key to understand its chemistry and to develop bio-mimetic catalysts.
We have recently determined the crystal structures of both MMOH bound to the activator (MMOB) and the inhibitor (MMOD), and structures revealed the key underlying molecular mechanisms.
In this proposed research, we aim to determine the EM structure of MMOH bound to the reductase (MMOR). Our preliminary data indicates that MMOH behaves well under the microscope. Since the crystal and NMR structures of both MMOH and MMOR are available, we can fit these known structures even in the intermediate resolution (~5Å). The structure will elucidate the atomic details of the mechanism by which MMOR shuttles electrons from NADH to MMOH. Additionally this work can suggest several potential conformational rearrangements that might occur in MMOH or MMOR during the reduction.

Project Details

Start Date
2020-02-15
End Date
2020-08-15
Status
Closed

Team

Principal Investigator

Uhn-soo Cho
Institution
University of Michigan

Team Members

Byung Chul Kim
Institution
University of Michigan