Genetic selection for and molecular dynamic modeling of a protein transmembrane domain multimerization motif from a random Escherichia coli genomic library

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Genetic selection for and molecular dynamic modeling of a protein transmembrane domain multimerization motif from a random Escherichia coli genomic library. / Leeds, J A; Boyd, D; Huber, D R; Sonoda, G K; Luu, H T; Engelman, D M; Beckwith, J.

In: Journal of Molecular Biology, Vol. 313, No. 1, 12.10.2001, p. 181-95.

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@article{82deb742c8784a12b4b455e75ab6fc71,
title = "Genetic selection for and molecular dynamic modeling of a protein transmembrane domain multimerization motif from a random Escherichia coli genomic library",
abstract = "In order to identify new transmembrane helix packing motifs in naturally occurring proteins, we have selected transmembrane domains from a library of random Escherichia coli genomic DNA fragments and screened them for homomultimerization via their abilities to dimerize the bacteriophage lambda cI repressor DNA-binding domain. Sequences were isolated using a modified lambda cI headpiece dimerization assay system, which was shown previously to measure transmembrane helix-helix association in the E. coli inner membrane. Screening resulted in the identification of several novel sequences that appear to mediate helix-helix interactions. One sequence, representing the predicted sixth transmembrane domain (TM6) of the E. coli protein YjiO, was chosen for further analysis. Using site-directed mutagenesis and molecular dynamics, a small set of models for YjiO TM6 multimerization interface interactions were generated. This work demonstrates the utility of combining in vivo genetic tools with computational systems for understanding membrane protein structure and assembly.",
keywords = "Genes, Bacterial, Viral Proteins, Genomic Library, Protein Subunits, Amino Acid Motifs, Repressor Proteins, Genetic Vectors, Escherichia coli, Cell Membrane, Molecular Sequence Data, Amino Acid Substitution, Bacteriophage lambda, Viral Regulatory and Accessory Proteins, Models, Molecular, Dimerization, DNA-Binding Proteins, Amino Acid Sequence, Recombinant Fusion Proteins, Protein Sorting Signals, Membrane Proteins, Protein Binding, Cloning, Molecular, Binding Sites, Protein Structure, Quaternary, Base Sequence, Escherichia coli Proteins, Protein Structure, Tertiary, Protein Transport",
author = "Leeds, {J A} and D Boyd and Huber, {D R} and Sonoda, {G K} and Luu, {H T} and Engelman, {D M} and J Beckwith",
note = "Copyright 2001 Academic Press.",
year = "2001",
month = oct,
day = "12",
doi = "10.1006/jmbi.2001.5007",
language = "English",
volume = "313",
pages = "181--95",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Elsevier",
number = "1",

}

RIS

TY - JOUR

T1 - Genetic selection for and molecular dynamic modeling of a protein transmembrane domain multimerization motif from a random Escherichia coli genomic library

AU - Leeds, J A

AU - Boyd, D

AU - Huber, D R

AU - Sonoda, G K

AU - Luu, H T

AU - Engelman, D M

AU - Beckwith, J

N1 - Copyright 2001 Academic Press.

PY - 2001/10/12

Y1 - 2001/10/12

N2 - In order to identify new transmembrane helix packing motifs in naturally occurring proteins, we have selected transmembrane domains from a library of random Escherichia coli genomic DNA fragments and screened them for homomultimerization via their abilities to dimerize the bacteriophage lambda cI repressor DNA-binding domain. Sequences were isolated using a modified lambda cI headpiece dimerization assay system, which was shown previously to measure transmembrane helix-helix association in the E. coli inner membrane. Screening resulted in the identification of several novel sequences that appear to mediate helix-helix interactions. One sequence, representing the predicted sixth transmembrane domain (TM6) of the E. coli protein YjiO, was chosen for further analysis. Using site-directed mutagenesis and molecular dynamics, a small set of models for YjiO TM6 multimerization interface interactions were generated. This work demonstrates the utility of combining in vivo genetic tools with computational systems for understanding membrane protein structure and assembly.

AB - In order to identify new transmembrane helix packing motifs in naturally occurring proteins, we have selected transmembrane domains from a library of random Escherichia coli genomic DNA fragments and screened them for homomultimerization via their abilities to dimerize the bacteriophage lambda cI repressor DNA-binding domain. Sequences were isolated using a modified lambda cI headpiece dimerization assay system, which was shown previously to measure transmembrane helix-helix association in the E. coli inner membrane. Screening resulted in the identification of several novel sequences that appear to mediate helix-helix interactions. One sequence, representing the predicted sixth transmembrane domain (TM6) of the E. coli protein YjiO, was chosen for further analysis. Using site-directed mutagenesis and molecular dynamics, a small set of models for YjiO TM6 multimerization interface interactions were generated. This work demonstrates the utility of combining in vivo genetic tools with computational systems for understanding membrane protein structure and assembly.

KW - Genes, Bacterial

KW - Viral Proteins

KW - Genomic Library

KW - Protein Subunits

KW - Amino Acid Motifs

KW - Repressor Proteins

KW - Genetic Vectors

KW - Escherichia coli

KW - Cell Membrane

KW - Molecular Sequence Data

KW - Amino Acid Substitution

KW - Bacteriophage lambda

KW - Viral Regulatory and Accessory Proteins

KW - Models, Molecular

KW - Dimerization

KW - DNA-Binding Proteins

KW - Amino Acid Sequence

KW - Recombinant Fusion Proteins

KW - Protein Sorting Signals

KW - Membrane Proteins

KW - Protein Binding

KW - Cloning, Molecular

KW - Binding Sites

KW - Protein Structure, Quaternary

KW - Base Sequence

KW - Escherichia coli Proteins

KW - Protein Structure, Tertiary

KW - Protein Transport

U2 - 10.1006/jmbi.2001.5007

DO - 10.1006/jmbi.2001.5007

M3 - Article

C2 - 11601855

VL - 313

SP - 181

EP - 195

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 1

ER -