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Determinants of translocation and folding of TreF, a trehalase of Escherichia coli.
J Biol Chem. 2000 Aug 04; 275(31):23439-45.JB

Abstract

One isoform of trehalase, TreF, is present in the cytoplasm and a second, TreA, in the periplasm. To study the questions of why one enzyme is exported efficiently and the other is not and whether these proteins can fold in their nonnative cellular compartment, we fused the signal sequence of periplasmic TreA to cytoplasmic TreF. Even though this TreF construct was exported efficiently to the periplasm, it was not active. It was insoluble and degraded by the periplasmic serine protease DegP. To determine why TreF was misfolded in the periplasm, we isolated and characterized Tre(+) revertants of periplasmic TreF. To further characterize periplasmic TreF, we used a genetic selection to isolate functional TreA-TreF hybrids, which were analyzed with respect to solubility and function. These data suggested that a domain located between residues 255 and 350 of TreF is sufficient to cause folding problems in the periplasm. In contrast to TreF, periplasmic TreA could fold into the active conformation in its nonnative cellular compartment, the cytoplasm, after removal of its signal sequence.

Authors+Show Affiliations

Fakultät für Biologie, Universität Konstanz, 78457 Konstanz, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10816581

Citation

Uhland, K, et al. "Determinants of Translocation and Folding of TreF, a Trehalase of Escherichia Coli." The Journal of Biological Chemistry, vol. 275, no. 31, 2000, pp. 23439-45.
Uhland K, Mondigler M, Spiess C, et al. Determinants of translocation and folding of TreF, a trehalase of Escherichia coli. J Biol Chem. 2000;275(31):23439-45.
Uhland, K., Mondigler, M., Spiess, C., Prinz, W., & Ehrmann, M. (2000). Determinants of translocation and folding of TreF, a trehalase of Escherichia coli. The Journal of Biological Chemistry, 275(31), 23439-45.
Uhland K, et al. Determinants of Translocation and Folding of TreF, a Trehalase of Escherichia Coli. J Biol Chem. 2000 Aug 4;275(31):23439-45. PubMed PMID: 10816581.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determinants of translocation and folding of TreF, a trehalase of Escherichia coli. AU - Uhland,K, AU - Mondigler,M, AU - Spiess,C, AU - Prinz,W, AU - Ehrmann,M, PY - 2000/5/19/pubmed PY - 2000/9/9/medline PY - 2000/5/19/entrez SP - 23439 EP - 45 JF - The Journal of biological chemistry JO - J Biol Chem VL - 275 IS - 31 N2 - One isoform of trehalase, TreF, is present in the cytoplasm and a second, TreA, in the periplasm. To study the questions of why one enzyme is exported efficiently and the other is not and whether these proteins can fold in their nonnative cellular compartment, we fused the signal sequence of periplasmic TreA to cytoplasmic TreF. Even though this TreF construct was exported efficiently to the periplasm, it was not active. It was insoluble and degraded by the periplasmic serine protease DegP. To determine why TreF was misfolded in the periplasm, we isolated and characterized Tre(+) revertants of periplasmic TreF. To further characterize periplasmic TreF, we used a genetic selection to isolate functional TreA-TreF hybrids, which were analyzed with respect to solubility and function. These data suggested that a domain located between residues 255 and 350 of TreF is sufficient to cause folding problems in the periplasm. In contrast to TreF, periplasmic TreA could fold into the active conformation in its nonnative cellular compartment, the cytoplasm, after removal of its signal sequence. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/10816581/Determinants_of_translocation_and_folding_of_TreF_a_trehalase_of_Escherichia_coli_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(19)65996-X DB - PRIME DP - Unbound Medicine ER -