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PMID: 9244262 Published · ppublish English Journal Article

Lipase secretion by bacterial hybrid ATP-binding cassette exporters: molecular recognition of the LipBCD, PrtDEF, and HasDEF exporters.

Journal of bacteriology ·Vol. 179 ·No. 15 ·1997-08-00 ·Pages 4754-60

Akatsuka H, Binet R, Kawai E, Wandersman C, Omori K

Abstract

Serratia marcescens secretes several proteins, such as the lipase LipA, the metalloprotease PrtA, and the heme-binding protein HasA, which is required for heme acquisition, through two N-terminal signal peptide-independent systems that are classified as bacterial ATP-binding cassette (ABC) exporters. One is the ABC exporter for HasA, consisting of the ABC protein HasD, the membrane fusion protein (MFP) HasE, and the outer membrane protein (OMP) HasF. The second, composed of LipB (an ABC protein), LipC (an MFP), and LipD (an OMP), promotes secretion of LipA and PrtA in Escherichia coli recombinant clones. PrtA, which shows homology to the Erwinia chrysanthemi metalloproteases, is efficiently secreted by E. coli cells carrying the E. chrysanthemi ABC exporter PrtD (ABC protein)-PrtE (MFP)-PrtF (OMP). The existence of distinct systems in this bacterium and of various substrates for these systems allowed the study of protein secretion by heterologous Has, Lip, and Prt systems and by Has-Lip and Lip-Prt hybrid exporters in the genuine host as well as in E. coli. For that purpose, lipB-, lipC-, and lipD-deficient mutants were isolated from S. marcescens 8000 and their secretion of LipA and PrtA was analyzed. This demonstrated that a unique exporter, the Lip apparatus, in S. marcescens secretes both LipA and PrtA. Hybrid exporters were tested for secretion of HasA and LipA. The LipB-HasE-HasF exporter allowed secretion of LipA but not HasA, showing that the ABC protein LipB is responsible for the substrate specificity. LipA, HasA, and E. chrysanthemi PrtC were secreted via heterologous exporters and via some hybrid exporters. Analysis of secretion via hybrid exporters showed that specific interactions occur between MFPs and OMPs in these systems. These genetic experiments demonstrated that specific interactions between the ABC protein and the MFP are required for the formation of active exporters.

MeSH Terms
ATP-Binding Cassette Transporters/genetics,metabolism Adenosine Triphosphatases Bacterial Outer Membrane Proteins/genetics,metabolism Bacterial Proteins/genetics,metabolism Carrier Proteins/genetics,metabolism Collagenases/genetics,metabolism Gene Deletion Genetic Complementation Test Heme-Binding Proteins Hemeproteins/genetics,metabolism Lipase/metabolism Membrane Proteins/genetics,metabolism Membrane Transport Proteins Metalloendopeptidases/genetics,metabolism Serratia marcescens/enzymology,genetics
Chemicals
ATP-Binding Cassette Transporters Bacterial Outer Membrane Proteins Bacterial Proteins Carrier Proteins HasE protein, Serratia marcescens HasF protein, bacteria Heme-Binding Proteins Hemeproteins LipA protein, Bacteria LipC protein, Serratia marcescens LipD protein, Serratia marcescens Membrane Proteins Membrane Transport Proteins PrtD protein, Erwinia chrysanthemi PrtF protein, bacteria Lipase Collagenases Metalloendopeptidases prtC protein, bacteria Adenosine Triphosphatases HasD protein, Serratia marcescens
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Akatsuka H
Lead Generation Research Laboratory, Tanabe Seiyaku Co., Ltd., Yodogawa-ku, Osaka, Japan.
Binet R
Kawai E
Wandersman C
Omori K
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.15.4754-4760.1997
Published
1997-08-00
Pages
4754-60
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179321
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.179.15.4754-4760.1997 PMC Full Text View journal details
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