Um mapa topográfico microscópico de complexidade da função celular: mero acaso, fortuita necessidade ou design inteligente?

quinta-feira, junho 13, 2019

Direct visualization of the E. coli Sec translocase engaging precursor proteins in lipid bilayers

Raghavendar Reddy Sanganna Gari1,*, Kanokporn Chattrakun1, Brendan P. Marsh1,†, Chunfeng Mao2, Nagaraju Chada1,‡, Linda L. Randall2 and Gavin M. King1,2,§

1Department of Physics and Astronomy, University of Missouri, Columbia, MO 65211, USA.

2Department of Biochemistry, University of Missouri, Columbia, MO 65211, USA.

↵§Corresponding author. Email:

↵* Present address: Department of Anesthesiology, Weill Cornell Medicine, New York, NY 10065, USA.

↵† Present address: Department of Applied Physics, Stanford University, Stanford, CA 94305, USA.

↵‡ Present address: Department of Biology, Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD 21218, USA.

Science Advances 12 Jun 2019: Vol. 5, no. 6, eaav9404

Fig. 6 Precursor-dependent model of translocase activity.


Escherichia coli exports proteins via a translocase comprising SecA and the translocon, SecYEG. Structural changes of active translocases underlie general secretory system function, yet directly visualizing dynamics has been challenging. We imaged active translocases in lipid bilayers as a function of precursor protein species, nucleotide species, and stage of translocation using atomic force microscopy (AFM). Starting from nearly identical initial states, SecA more readily dissociated from SecYEG when engaged with the precursor of outer membrane protein A as compared to the precursor of galactose-binding protein. For the SecA that remained bound to the translocon, the quaternary structure varied with nucleotide, populating SecA2 primarily with adenosine diphosphate (ADP) and adenosine triphosphate, and the SecA monomer with the transition state analog ADP-AlF3. Conformations of translocases exhibited precursor-dependent differences on the AFM imaging time scale. The data, acquired under near-native conditions, suggest that the translocation process varies with precursor species.

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