PMID- 9352918
OWN - NLM
STAT- MEDLINE
DCOM- 19971120
LR  - 20190508
IS  - 0021-9193 (Print)
IS  - 1098-5530 (Electronic)
IS  - 0021-9193 (Linking)
VI  - 179
IP  - 21
DP  - 1997 Nov
TI  - Topological analysis of the membrane-bound glucosyltransferase, MdoH, required 
      for osmoregulated periplasmic glucan synthesis in Escherichia coli.
PG  - 6692-8
AB  - The MdoH protein is essential for synthesis of the osmoregulated periplasmic 
      glucans, known as membrane-derived oligosaccharides (MDOs), in Escherichia coli. 
      Mutants lacking MdoH are deficient in a glucosyltransferase activity assayed in 
      vitro. The MdoH protein is the product of the second gene of an operon, and it 
      has been shown to span the cytoplasmic membrane. The MdoH protein comprises 847 
      amino acids and is poorly expressed as observed by sodium dodecyl 
      sulfate-polyacrylamide gel electrophoresis. We have experimentally measured the 
      topological organization of MdoH within the membrane by construction of fusions 
      to beta-lactamase as a reporter. Analysis of 51 different MdoH-beta-lactamase 
      fusions suggested that the MdoH protein crosses the cytoplasmic membrane eight 
      times, with the N and C termini in the cytoplasm. Moreover, a 310-amino-acid 
      domain is present in the cytoplasm between the second and third transmembrane 
      segments. It was deduced from the measurement of the MDO biosynthetic activity of 
      truncated or fused MdoH proteins that almost all the C-terminal residues are 
      necessary for this activity. The model of the MdoH protein in the membrane 
      suggests that this protein could be directly involved in the translocation of 
      nascent polyglucose chains to the periplasmic space.
FAU - Debarbieux, L
AU  - Debarbieux L
AD  - Laboratoire de Chimie Biologique, UMR111 du CNRS, Universite des Sciences et 
      Technologies de Lille, Villeneuve d'Ascq, France.
FAU - Bohin, A
AU  - Bohin A
FAU - Bohin, J P
AU  - Bohin JP
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Bacteriol
JT  - Journal of bacteriology
JID - 2985120R
RN  - 0 (Bacterial Proteins)
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Glucans)
RN  - 0 (Membrane Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (opgH protein, E coli)
RN  - EC 2.4.1.- (Glucosyltransferases)
RN  - EC 3.4.21.64 (Endopeptidase K)
RN  - EC 3.5.2.6 (beta-Lactamases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Ampicillin Resistance
MH  - Bacterial Proteins/*chemistry/genetics
MH  - Cell Compartmentation
MH  - Endopeptidase K/metabolism
MH  - Escherichia coli/metabolism
MH  - *Escherichia coli Proteins
MH  - Glucans
MH  - Glucosyltransferases/*chemistry/genetics
MH  - Membrane Proteins/*chemistry/genetics
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Periplasm
MH  - Protein Conformation
MH  - Recombinant Fusion Proteins
MH  - Sequence Deletion
MH  - Water-Electrolyte Balance
MH  - beta-Lactamases/genetics
PMC - PMC179597
EDAT- 1997/11/14 00:00
MHDA- 1997/11/14 00:01
PMCR- 1997/11/01
CRDT- 1997/11/14 00:00
PHST- 1997/11/14 00:00 [pubmed]
PHST- 1997/11/14 00:01 [medline]
PHST- 1997/11/14 00:00 [entrez]
PHST- 1997/11/01 00:00 [pmc-release]
AID - 10.1128/jb.179.21.6692-6698.1997 [doi]
PST - ppublish
SO  - J Bacteriol. 1997 Nov;179(21):6692-8. doi: 10.1128/jb.179.21.6692-6698.1997.