Escherichia coli K-12 substr. MG1655 Polypeptide: periplasmic chaperone, effects translocation of lipoproteins from inner membrane to outer

Gene: lolA Accession Numbers: G6465 (EcoCyc), b0891, ECK0882

Synonyms: yzzV, lplA, p20

Regulation Summary Diagram: ?

Regulation summary diagram for lolA

Lipoproteins contain a fatty acyl chain covalently attached to the N-terminal cysteine residue. The amino acids at positions 2 and 3 after the cysteine residue function as a sorting signal; generally, aspartate at +2 causes the lipoprotein to be retained in the inner membrane [Seydel99, Terada01].

The LolA protein was initially identified as essential for the translocation of the major outer membrane lipoprotein Lpp from the inner to the outer membrane [Matsuyama95]. In vitro, translocation requires NTP hydrolysis and the presence of the outer membrane sorting signal [Yakushi98]. All lipoproteins with an outer membrane localization signal form a complex with LolA; efficient release from the inner membrane and incorporation of the lipoprotein into the outer membrane requires formation of the LolA-lipoprotein complex and the presence of LolB in the outer membrane [Yokota99] as well as aminoacylation of the protein [Fukuda02].

LolA is essential for growth [Tajima98]. Site-directed mutagenesis has identified Arg43 as the amino acid involved in the transfer of lipoprotein to LolB [Miyamoto01]. An F47E mutant has a dominant negative effect by remaining tightly associated with the LolCDE transporter [Miyamoto02]. Overexpression of lolA activates the RcsCDB Two-Component Signal Transduction System, causing the production of an extracellular polysaccharide capsule of colanic acid [Chen01c]. Mutations which cause the accumulation of outer membrane specific lipoproteins in the inner membrane induce expression of lolA via the Rcs phosphorelay system [Tao12a].

A crystal structure of LolA has been solved at 1.65 Å resolution. An 11-stranded anti-parallel β-sheet forms a β-barrel structure with three α-helices constituting a 'lid' which is believed to open and close upon accomodation and release of lipoproteins [Takeda03, Oguchi08]. The inner surface of the β-sheet and α-helices consists of hydrophobic residues and forms a cavity which binds to the acyl chains of lipoproteins [Okuda09]. In vivo photo cross-linking between LolA and mLolB suggests that the external surface of the mLolB hydrophobic cavity interacts with the internal surface of the LolA hydrophobic cavity and a 'mouth-to-mouth' model for the transfer of lipoproteins has been proposed [Okuda09].

Reviews: [Narita04, Tokuda04]

Locations: periplasmic space

Map Position: [936,592 -> 937,206] (20.19 centisomes, 73°)
Length: 615 bp / 204 aa

Molecular Weight of Polypeptide: 22.628 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0003031 , DIP:DIP-35675N , EchoBASE:EB2548 , EcoGene:EG12684 , EcoliWiki:b0891 , Mint:MINT-1249033 , ModBase:P61316 , OU-Microarray:b0891 , PortEco:lolA , PR:PRO_000023103 , Pride:P61316 , Protein Model Portal:P61316 , RefSeq:NP_415411 , RegulonDB:G6465 , SMR:P61316 , String:511145.b0891 , UniProt:P61316

Relationship Links: InterPro:IN-FAMILY:IPR004564 , InterPro:IN-FAMILY:IPR018323 , InterPro:IN-FAMILY:IPR029046 , PDB:Structure:1IWL , PDB:Structure:1UA8 , PDB:Structure:2ZPC , PDB:Structure:2ZPD , PDB:Structure:3KSN , Pfam:IN-FAMILY:PF03548

Genetic Regulation Schematic: ?

Genetic regulation schematic for lolA

GO Terms:

Biological Process: GO:0042953 - lipoprotein transport Inferred from experiment Inferred by computational analysis [GOA01, Matsuyama95]
GO:0044874 - lipoprotein localization to outer membrane Inferred from experiment [Matsuyama95]
GO:0072323 - chaperone-mediated protein transport across periplasmic space Inferred from experiment [Matsuyama95]
GO:0006810 - transport Inferred by computational analysis [UniProtGOA11]
GO:0015031 - protein transport Inferred by computational analysis [UniProtGOA11, GOA06]
Molecular Function: GO:0042954 - lipoprotein transporter activity Inferred from experiment Inferred by computational analysis [GOA01, Matsuyama95]
GO:0008565 - protein transporter activity Inferred by computational analysis [GOA06]
Cellular Component: GO:0030288 - outer membrane-bounded periplasmic space Inferred from experiment Inferred by computational analysis [DiazMejia09, Han14, LopezCampistrou05, Matsuyama95]
GO:0042597 - periplasmic space Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA06, GOA01]

MultiFun Terms: transport Putative uncharacterized transport protein

Essentiality data for lolA knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox No 37 Aerobic 7   No [Baba06, Comment 2]

Sequence Features

Protein sequence of periplasmic chaperone, effects translocation of lipoproteins from inner membrane to outer with features indicated

Feature Class Location Common Name Citations Comment
Signal-Sequence 1 -> 22 signal peptide
Alpha-Helix-Region 23 -> 34 α1
Mutagenesis-Variant 64  
[Miyamoto01, UniProt11]
UniProt: Loss of ability to transfer lipoproteins to lolB.
Mutagenesis-Variant 68  
[Miyamoto02, UniProt11]
UniProt: Loss of ability to bind lipoproteins.
Beta-Strand-Region 78 -> 84 β4
Beta-Strand-Region 87 -> 92 β5
Beta-Strand-Region 96 -> 102 β6
Alpha-Helix-Region 110 -> 117 α2
Alpha-Helix-Region 119 -> 124 α3
Beta-Strand-Region 126 -> 131 β7
Beta-Strand-Region 133 -> 139 β8
Beta-Strand-Region 147 -> 154 β9
Beta-Strand-Region 158 -> 165 β10
Beta-Strand-Region 170 -> 182 β11

Gene Local Context (not to scale): ?

Gene local context diagram

Transcription Unit:

Transcription-unit diagram


Markus Krummenacker on Tue Oct 14, 1997:
Gene object created from Blattner lab Genbank (v. M52) entry.


Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Chen01c: Chen MH, Takeda S, Yamada H, Ishii Y, Yamashino T, Mizuno T (2001). "Characterization of the RcsC-->YojN-->RcsB phosphorelay signaling pathway involved in capsular synthesis in Escherichia coli." Biosci Biotechnol Biochem 65(10);2364-7. PMID: 11758943

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Fukuda02: Fukuda A, Matsuyama S, Hara T, Nakayama J, Nagasawa H, Tokuda H (2002). "Aminoacylation of the N-terminal cysteine is essential for Lol-dependent release of lipoproteins from membranes but does not depend on lipoprotein sorting signals." J Biol Chem 277(45);43512-8. PMID: 12198129

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Han14: Han MJ, Kim JY, Kim JA (2014). "Comparison of the large-scale periplasmic proteomes of the Escherichia coli K-12 and B strains." J Biosci Bioeng 117(4);437-42. PMID: 24140104

Link97: Link AJ, Robison K, Church GM (1997). "Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12." Electrophoresis 18(8);1259-313. PMID: 9298646

LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

Matsuyama95: Matsuyama S, Tajima T, Tokuda H (1995). "A novel periplasmic carrier protein involved in the sorting and transport of Escherichia coli lipoproteins destined for the outer membrane." EMBO J 14(14);3365-72. PMID: 7628437

Miyamoto01: Miyamoto A, Matsuyama Si , Tokuda H (2001). "Mutant of LolA, a lipoprotein-specific molecular chaperone of Escherichia coli, defective in the transfer of lipoproteins to LolB." Biochem Biophys Res Commun 287(5);1125-8. PMID: 11587539

Miyamoto02: Miyamoto A, Matsuyama S, Tokuda H (2002). "Dominant negative mutant of a lipoprotein-specific molecular chaperone, LolA, tightly associates with LolCDE." FEBS Lett 528(1-3);193-6. PMID: 12297303

Narita04: Narita S, Matsuyama S, Tokuda H (2004). "Lipoprotein trafficking in Escherichia coli." Arch Microbiol 182(1);1-6. PMID: 15221203

Oguchi08: Oguchi Y, Takeda K, Watanabe S, Yokota N, Miki K, Tokuda H (2008). "Opening and closing of the hydrophobic cavity of LolA coupled to lipoprotein binding and release." J Biol Chem. PMID: 18617521

Okuda09: Okuda S, Tokuda H (2009). "Model of mouth-to-mouth transfer of bacterial lipoproteins through inner membrane LolC, periplasmic LolA, and outer membrane LolB." Proc Natl Acad Sci U S A 106(14);5877-82. PMID: 19307584

Seydel99: Seydel A, Gounon P, Pugsley AP (1999). "Testing the '+2 rule' for lipoprotein sorting in the Escherichia coli cell envelope with a new genetic selection." Mol Microbiol 34(4);810-21. PMID: 10564520

Tajima98: Tajima T, Yokota N, Matsuyama S, Tokuda H (1998). "Genetic analyses of the in vivo function of LolA, a periplasmic chaperone involved in the outer membrane localization of Escherichia coli lipoproteins." FEBS Lett 439(1-2);51-4. PMID: 9849875

Takeda03: Takeda K, Miyatake H, Yokota N, Matsuyama S, Tokuda H, Miki K (2003). "Crystal structures of bacterial lipoprotein localization factors, LolA and LolB." EMBO J 22(13);3199-209. PMID: 12839983

Tao12a: Tao K, Narita S, Tokuda H (2012). "Defective lipoprotein sorting induces lolA expression through the Rcs stress response phosphorelay system." J Bacteriol 194(14);3643-50. PMID: 22563052

Terada01: Terada M, Kuroda T, Matsuyama SI, Tokuda H (2001). "Lipoprotein sorting signals evaluated as the LolA-dependent release of lipoproteins from the cytoplasmic membrane of Escherichia coli." J Biol Chem 276(50);47690-4. PMID: 11592971

Tokuda04: Tokuda H, Matsuyama S (2004). "Sorting of lipoproteins to the outer membrane in E. coli." Biochim Biophys Acta 1693(1);5-13. PMID: 15276320

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

Yakushi98: Yakushi T, Yokota N, Matsuyama S, Tokuda H (1998). "LolA-dependent release of a lipid-modified protein from the inner membrane of Escherichia coli requires nucleoside triphosphate." J Biol Chem 273(49);32576-81. PMID: 9829994

Yokota99: Yokota N, Kuroda T, Matsuyama S, Tokuda H (1999). "Characterization of the LolA-LolB system as the general lipoprotein localization mechanism of Escherichia coli." J Biol Chem 274(43);30995-9. PMID: 10521496

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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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