Molecular characterization of PulE, a protein required for pullulanase secretion
- 1 April 1994
- journal article
- Published by Wiley in Molecular Microbiology
- Vol. 12 (2) , 287-299
- https://doi.org/10.1111/j.1365-2958.1994.tb01017.x
Abstract
Summary: pulE, one of 14 genes specifically required for pullulanase secretion in Klebsiella oxytoca, codes for a putative nucleotide‐binding protein. Subcellular fractioNatlon indicated that the majority of PulE in Escherichia coli cells expressing all 14 secretion genes is mainly associated with the cytoplasmic membrane through both hydrophobic and non‐hydrophobic interactions. Mutational analysis revealed that one of the two regions of PulE that are conserved in many nucteotide‐binding proteins (Walker box A) is essential for pullulanase secretion. Likewise, mutations that removed aspartate residues from each of two regions immediately downstream from the Walker box A also reduced secretion. These aspartate‐rich regions are highly conserved in all 16 known PulE homologues but not in any other nucleotide‐binding proteins. Altogether, these results Indicate that PulE might belong to a new family of nucleotide‐binding proteins. The protein could not be cross‐linked to the photoactivatable ATP analogue azido‐ATP, however. Most pulE point or deletion mutations which prevented pullulanase secretion exhibited transdominance when expressed at high levels in cells producing wild‐type PulE protein. Evidence presented suggests that PulE might be a homodimer.Keywords
This publication has 44 references indexed in Scilit:
- The Rieske protein from Paracoccus denitrificans is inserted into the cytoplasmic membrane by the twin‐arginine translocaseThe FEBS Journal, 2006
- Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein‐secretion apparatus: a general system for the formation of surface‐associated protein complexesMolecular Microbiology, 1993
- Processing and methylation of PulG, a pilin‐like component of the general secretory pathway of Klebsiella oxytocaMolecular Microbiology, 1993
- ABC Transporters: From Microorganisms to ManAnnual Review of Cell Biology, 1992
- The Aeromonas hydrophila exeE gene, required both for protein secretion and normal outer membrane biogenesis, is a member of a general secretion pathwayMolecular Microbiology, 1992
- Pullulanase secretion in Escherichia coli K-12 requires a cytoplasmic protein and a putative polytopic cytoplasmic membrane proteinMolecular Microbiology, 1992
- Characterisation of a Pseudomonas aeruginosa twitching motility gene and evidence for a specialised protein export system widespread in eubacteriaGene, 1991
- Two distinct steps in pullulanase secretion by Escherichia coli K12Molecular Microbiology, 1991
- The general protein‐export pathway is directly required for extracellular pullulanase secretion in Escherichia coli k12Molecular Microbiology, 1991
- Kanamycin-resistant vectors that are analogues of plasmids pUC8, pUC9, pEMBL8 and pEMBL9Gene, 1986