期刊
出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.2018122118
关键词
electrochromic shift; twin arginine translocon; Sec; protein translocation; toroidal pore
资金
- Division of Chemical Sciences, Geosciences, and Biosciences, Office of Basic Energy Sciences of the US Department of Energy [DE-SC0020304]
- U.S. Department of Energy (DOE) [DE-SC0020304] Funding Source: U.S. Department of Energy (DOE)
The mechanism and pore architecture of the Tat complex during transport of folded substrates remain a mystery, while the toroidal pore model better accounts for the high-energy barrier in Tat transport. The proteinaceous SecY pore undergoes conformational shifts between closed and opened states for translocating unfolded substrate chains.
The mechanism and pore architecture of the Tat complex during transport of folded substrates remain a mystery, partly due to rapid dissociation after translocation. In contrast, the proteinaceous SecY pore is a persistent structure that needs only to undergo conformational shifts between closed and opened states when translocating unfolded substrate chains. Where the proteinaceous pore model describes the SecY pore well, the toroidal pore model better accounts for the high-energy barrier that must be overcome when transporting a folded substrate through the hydrophobic bilayer in Tat transport. Membrane conductance behavior can, in principle, be used to distinguish between toroidal and proteinaceous pores, as illustrated in the examination of many antimicrobial peptides as well as mitochondrial Bax and Bid. Here, we measure the electrochromic shift (ECS) decay as a proxy for conductance in isolated thylakoids, both during protein transport and with constitutively assembled translocons. We find that membranes with the constitutively assembled Tat complex and those undergoing Tat transport display conductance characteristics similar to those of resting membranes. Membranes undergoing Sec transport and those with the substrate-engaged SecY pore result in significantly more rapid electric field decay. The responsiveness of the ECS signal in membranes with active SecY recalls the steep relationship between applied voltage and conductance in a proteinaceous pore, while the nonaccelerated electric field decay with both Tat transport and the constitutive Tat complex under the same electric field is consistent with the behavior of a toroidal pore.
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