Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.3.1.269 extracted from

  • Nozeret, K.; Boucharlat, A.; Agou, F.; Buddelmeijer, N.
    A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein apolipoprotein N-acyltransferase (Lnt) (2019), Sci. Rep., 9, 15978 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.3.1.269

Cloned(Commentary)

Cloned (Comment) Organism
gene lnt, recombinant overexpression of Strep-tagged wild-type and mutant enzymes in Escherichia coli Escherichia coli

Protein Variants

Protein Variants Comment Organism
C387S site-directed mutagenesis, the mutant catalyzes the 1st step of the N-acyl transfer reaction and forms a oxygen-ester acyl intermediate but is unable to transfer the acyl group onto apolipoprotein Escherichia coli
K335A site-directed mutagenesis, inactive mutant Escherichia coli

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane integral membrane protein Escherichia coli 16020
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
a phosphoglycerolipid + an [apolipoprotein]-S-1,2-diacyl-sn-glyceryl-L-cysteine Escherichia coli
-
 a 1-lyso-phosphoglycerolipid + a [lipoprotein]-N-acyl-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P23930
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli by affinity chromatography and gel filtration Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-alkyne-2-acyl-phosphatidylethanolamine + [FSL-1 apolipopeptide]-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 Escherichia coli 1-lyso-2-acyl-phosphatidylethanolamine + [FSL-1 lipopeptide]-N-alkyne-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 ?
1-palmitoyl alkyne-2-oleoyl-sn-glycero-3-phosphatidylethanolamine + [FSL-1 apolipopeptide]-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 Escherichia coli 1-lyso-2-oleoyl-sn-glycero-3-phosphatidylethanolamine + [FSL-1 lipopeptide]-N-palmitoyl alkyne-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 ?
a phosphoglycerolipid + an [apolipoprotein]-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 Escherichia coli a 1-lyso-phosphoglycerolipid + a [lipoprotein]-N-acyl-S-1,2-diacyl-sn-glyceryl-L-cysteine
-
 ?
additional information enzyme Lnt uses apolipoprotein (S-diacylglyceryl protein) as protein substrate in the reaction. Lnt catalyzes a two-step reaction via a ping-pong mechanism, whereby in the first step, a stable thioester acyl-enzyme intermediate is formed upon hydrolysis of phospholipid. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylethanolamine (POPE) is the preferred substrate for Lnt. When the lysophospholipid by-product is released, N-acyl transfer onto apolipoprotein occurs in the second step of the reaction. The enzyme is unable to catalyze the N-acyl transferase reaction in the presence of POPE and no effect is observed with PE-biotin. Various FSL-1 peptide substrates are N-acylated by Lnt in vitro. N-acyltransferase activity of Lnt is monitored as a shift in migration of the lipopeptide FSL-1 conjugated either with biotin or fluorescein, and detection of Lnt activity by fluorescence spectroscopy, method evaluation and optimization, overview Escherichia coli ?
-
-

Synonyms

Synonyms Comment Organism
lnt
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
 assay at Escherichia coli