Title: Thylakoid-integrated recombinant Hcf106 participates in the chloroplast twin arginine transport system

Journal Article · · Plant Direct
DOI: https://doi.org/10.1002/pld3.90 · OSTI ID:1593749
 [1];  [2];  [3]; ORCiD logo [3]
  1. Miami Univ., Oxford, OH (United States); Johns Hopkins Univ., Baltimore, MD (United States); Miami University (OH)
  2. Miami Univ., Oxford, OH (United States); Wright State Univ., Dayton, OH (United States)
  3. Miami Univ., Oxford, OH (United States)
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The chloroplast twin arginine transport (cpTat) system distinguishes itself as a protein transport pathway by translocating fully folded proteins, using the proton-motive force (PMF) as the sole source of energy. The cpTat pathway is evolutionarily conserved with the Tat pathway found in the plasma membrane of many prokaryotes. The cpTat (Escherichia coli) system uses three proteins, Tha4 (TatA),Hcf106 (TatB), and cpTatC (TatC), to form a transient translocase allowing the pas-sage of precursor proteins. Briefly, cpTatC and Hcf106, with Tha4, form the initial receptor complex responsible for precursor protein recognition and binding in an energy-independent manner, while a separate pool of Tha4 assembles with the pre-cursor-bound receptor complex in the presence the PMF. Analysis by blue-native polyacrylamide gel electrophoresis (BN-PAGE) shows that the receptor complex, in the absence of precursor, migrates near 700 kDa and contains cpTatC and Hcf106with little Tha4 remaining after detergent solubilization. To investigate the role thatHcf106 may play in receptor complex oligomerization and/or stability, systematic cysteine substitutions were made in positions from the N-terminal transmembrane domain to the end of the predicted amphipathic helix of the protein. BN-PAGE analysis allowed us to identify the locations of amino acids in Hcf106 that were critical for interacting with cpTatC. Oxidative cross-linking allowed us to map interactions of the transmembrane domain and amphipathic helix region of Hcf106. In addition, we showed that in vitro expressed, integrated Hcf106 can interact with the precursor signal peptide domain and imported cpTatC, strongly suggesting that a subpopulation of the integrated Hcf106 is participating in competent cpTat complexes.

Research Organization:
Miami Univ., Oxford, OH (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division
Grant/Contract Number:
SC0014441
OSTI ID:
1593749
Journal Information:
Plant Direct, Journal Name: Plant Direct Journal Issue: 10 Vol. 2; ISSN 2475-4455
Publisher:
Wiley and American Society of Plant Biologists and Society for Experimental BiologyCopyright Statement
Country of Publication:
United States
Language:
English

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59 BASIC BIOLOGICAL SCIENCES
cross‐linking
oligomerization
receptor complex
truncation
twin arginine transport