Information on EC 2.3.1.51 - 1-acylglycerol-3-phosphate O-acyltransferase

C4B4E7

low expression level

Q8GXU8, Q8L4Y2, Q8LG50, Q9LHN4, Q9SYC8

the endoplasmic reticulum-located LPAT2 is essential for female but not male gametophyte development in Arabidopsis thaliana

gut

mAGPAT1, mAGPAT2, mAGPAT3, mAGPAT4 is expressed at low levels, mAGPAT5 expressed at intermediate levels

expression of AGPAT7

CD4+

lactating

Oryctolagus cuniculus

pancreatic cancer cell

Q8GXU8, Q8L4Y2, Q8LG50, Q9LHN4, Q9SYC8

LPAT2 predominantly

etiolated

C4B4E7

testicular cell line

expression of AGPAT7

physiological significance analyzed in white adipose tissue of mice, soluble lysophosphatidic acid acyltransferase activity, existence of an alternate cytosolic phosphatidic acid biosynthetic pathway

additional information

16 entries

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

chloroplast

mature protein of 32000 Da processed from the precursor protein of 38000 Da

LPAAT2

isoform AGPAT1 localizes to cytoskeletal components, particularly to areas of actin polymerization in myoblasts

endoplasmic reticulum

13 entries

Golgi membrane

mitochondrial outer membrane

integral enzyme

139

outer mitochondrial membrane

mitochondrion

6 entries

sarcoplasmic reticulum

Oryctolagus cuniculus

additional information

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

5a4h, download, 3D-view

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

29000

apparent, gel filtration

32000

34800

calculated from amino acid sequence

37000

38000

predicted, protein encoded by the BAT2 cDNA

487207

43300

C4B4E7

x * 43300, about, sequence calculation, x * 37000, about, recombinant FLAg-tagged enzyme, SDS-PAGE

705005

43380

Q9NRZ7, Q9NUQ2

x * 43380, claculated; x * 43380, claculated

720178

43723

Sterculia foetida

V9TK35

x * 43723, calculated from amino acid sequence

734968

43800

x * 43800, calculated from amino acid sequence

44000

50000

x * 50000, SDS-PAGE

61180

P33333, Q08548

partially purified Slc1p, SDS-PAGE, Western Blot

687624

additional information

45 kDa protein in Western Blot and mass spectrometry analysis, subcellular fractions of white adipose tissue

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

8 entries

additional information

Q9DBL9

the CGI-58 carboxyl terminus includes a highly conserved consensus sequence, HXXXXD, required for acyltransferase activity

705011

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

6.5 - 7.5

lowering the assay pH from 7.5 to 6.5 results in a loss of approximately 20% of the enzyme activity

487198

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

t1/2: 62 min

10 min, 60% loss of activity

705011

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

ethylene glycol, 20% v/v, stabilizes

Rattus norvegicus

486486, 486634

slow freezing inactivates

the long time required for fractionation after hypotonic hemolysis probably contributes to the loss of activity

Sus scrofa

487192

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

-15°C to 4°C, particulate preparation, fast-frozen in liquid N2, unstable

-15°C, less than a day

-20°C, microsomal preparation, at least 3 weeks

Spinacia oleracea

487188

-20°C, particulate preparation, several days

Pisum sativum

487184

-80°C, particulate preparation, fast-frozen in liquid N2, 3 months

4°C, high-speed supernatant preparation, 50% loss of activity, overnight

487198

4°C, high-speed supernatant, reactivated with phospholipids, no loss of activity

487198

4°C, soluble preparation, at least 1 day

Pisum sativum

487184

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

487179

gel filtration and SDS-PAGE, recombinant protein

687813

gel filtration and SDS-PAGE, recombinant protein, mutant and wild-type

gel filtration, wild-type and mutant protein

identification of enzyme using solubilization with CHAPS and column chromatography on Red 120 agarose, Bio-Rad HA HT, 12:0-CoA Sepharose 4B, and Superose 12

native enzyme partially by microsome preparation

Linum usitatissimum

675752

partial

3 entries

partial, separated from EC 2.3.1.15 using Triton X-100, the enzyme is not solubilized by Triton X-100 or cholate

487180

partial; using solubilization with CHAPS at high ionic strength, and anion-exchange chromatography on Mono Q column

Pisum sativum

487184

partial; using solubilization with Triton X-100 and gel filtration, separation from EC 2.3.1.23

Rattus norvegicus

486634

partially purified, gel filtration; partially purified, gel filtration

P33333, Q08548

687624

recombinant enzyme partially from CHO-K1 cell microsomes

O15120

674530

recombinant His-tagged At4g24160 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

657854

AGPAT10/GPAT3, expression in CHO cells endoplasmic reticulum, and as AGPAT-active enzyme in HEK-293 cells via transfection with an adenoviral vector, overexpression of the AGPAT-active enzyme in Huh-7 cells depleted of endogenous AGPAT activity by shRNA-AGPAT2-lentivirus

705206

amino acid sequence data used to isolate a full-length clone from a coconut endosperm cDNA library, expression of the cDNA in Escherichia coli

expressed in Arabidopsis thaliana

Sterculia foetida

V9TK35

734968

expressed in Baculovirus-Sf9 insect cells, pESC-URA yeast expression vector with a c-myc-epitope, transformation into Escherichia coli after site-directed mutagenesis

expressed in CHO-K1 cells

735599

expressed in Escherichia coli strain HB101, transformed into Saccharomyces cerevisiae, strains, plasmids and sources of parent vectors listed; expressed in Escherichia coli strain HB101, transformed into Saccharomyces cerevisiae, strains, plasmids and sources of parent vectors listed

P33333, Q08548

687624

expressed in Escherichia coli, overexpression in Saccharomyces cerevisiae, expression plasmid pPS189

687813

expressed in Escherichia coli, plasmids pCR2.1, pDrive, pcDNA 3.1(C)neo, construction of the wild-type fusion protein with GFP, overexpression in CHO-cells, coexpression in CHO-cells with AGPAT1

Q8NF37

688005

expressed in HEK-293 cells

Q3TFD2

736479

expressed in HEK-293 or Sf9 insect cells

735714

expressed in Saccharomyces cerevisiae strain INVSc1 and Nicotiana tabacum seeds

Q42670

737242

expression in C2C12 cell

O35083

719337

expression in Chineses hamster ovary cells

expression in CHO cell, HeLa cell and HEK-293 cell

Q7L5N7

720175

expression in Escherichia coli

expression in Escherichia coli JC201, a mutant strain deficient in the enzyme activity

487202

expression in Escherichia coli; expression of active His6-tagged CGI-58 in Escherichia coli

Q9DBL9

705011

expression in HEK-293 cell; expression in HEK-293 cell

Q9NRZ7, Q9NUQ2

720178

expression in insect and mammalian cell lines. The enzyme gene is located in the MHC class III region

487204

expression in JC201 Escherichia coli cells

487210

expression in osteosarcoma cell

720813

expression of a cDNA encoding protein with enzyme activity in transgenic plants. Expression in canola results in incorporation of laurate into the sn-2 position of triacylglycerol

487206

expression via recombinant adenoviruses in Agpat2 -/- mice; expression via recombinant adenoviruses in Agpat2 -/- mice

719977

gene AGPAT2, expression of C-terminally HA-tagged AGPAT2 in CHO-K1 cells

O15120

674530

gene AGPAT7, DNA and amino acid sequence determination and analysis, chromosomal mapping to 15q14, expression analysis

Q643R3

673280

gene AGPAT8, DNA and amino acid sequence determination and analysis, localization on chromosome 17, genetic structure, expression analysis

671597

gene AGPAT8, DNA and amino acid sequence determination and analysis, localization on chromosome 2p23.1, genetic structure, expression analysis, stable expression of wild-type and mutant enzymes in CHO cells

671597

gene At4g24160, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3)

Arabidopsis thaliana

O22975

706324

gene LPAT1, genetic structure, phylogenetic tree; gene LPAT2, genetic structure, LPAT2-cDNA driven by an LPAT2 promoter functionally complements insertion mutant lpat2 in transformed heterozygous mutants to produce the lpat2/lpat2 genotype, overview, phylogenetic tree, functional expression in Saccharomyces cerevisiae, genotyping; gene LPAT3, genetic structure, phylogenetic tree, functional expression in Saccharomyces cerevisiae, genotyping; gene LPAT4, genetic structure, phylogenetic tree, expression in Saccharomyces cerevisiae results in an inactive enzyme; gene LPAT5, genetic structure, phylogenetic tree, expression in Saccharomyces cerevisiae results in an inactive enzyme

Arabidopsis thaliana

Q8GXU8, Q8L4Y2, Q8LG50, Q9LHN4, Q9SYC8

676410

gene plsC, subcloning in Escherichia coli strain DH5alpha

Sinorhizobium meliloti

704819

genes BAT1.13 and BAT1.5, cloning of two cDNAs encoding microsomal isozymes from an immature embryo library, sequence comparisons, functional expression of the two rapeseed microsomal LPAAT isozymes in Arabidopsis thaliana, using Agrobacterium tumefaciens strain C58C1 transfection, leading to enhanced oil content and seed weight, tissue localization study, overview. Complementationof a thermosensitive Escherichia coli strain JC201 by expression of Bat1.5 and BAT1.13, overview

706333

isolation and characterization of a gonococcal mutant

Neisseria gonorrhoeae

Q59601

487200

isolation and characterization of a meningococcal mutant, cloning of the meningococcal mutant gene

Neisseria meningitidis

Q59618

487200

isolation of a cDNA encoding an enzyme by functional complementation of the Escherichia coli mutant plsC with an immature embryo cDNA library of oilseed rape. Transformation of the acyltransferase-deficient Escherichia coli strain JC201 with the cDNA sequence BAT2 alleviates the temperature-sensitive phenotype of the plsC mutant and confers a palmitoyl-CoA-preferring enzyme activity to membrane fractions. Mapping of the BAT2 genes and sequence analysis of the BAT2 cDNA clone

487207

isolation of a potential cDNA for the enzyme by heterologous screening with the maize pMAT1 clone, clone pLAT1; isolation of a potential cDNA for the enzyme by in vivo complementation of the Escherichia coli mutant JC201, clone pLAT2 is an enzyme clone

Limnanthes douglasii

Q40119, Q42870

487199

isolation of the JC201-plsC gene fragment containing the entire sequence of the enzyme. Complementation of the JC201 phenotype: transformation of JC201 cells with pplSC6 or pplsC6.3. Transformation of XL1-Blue cells with the control plasmid pplsC6.3, and the experimental plasmids pplsC6.3A or pplsC6.3L

487205

LPAAT3, phylogenetic analysis, expression of the FLAG-tagged enzyme in CHO-K1 cells

C4B4E7

705005

modification of the gene of Escherichia coli which encodes the enzyme by inserting an endoplasmic reticulum retrieval signal to its 3'end and introduccion into rapeseed plants under the control of a napin promotor

487203

overexpressed in Escherichia coli BL21 (DE3), pCMV-SPORT6 vector, overexpressed in Saccharomyces cerevisiae, pYES2 vector

stable functional expression of GFP-tagged LPAAT3 in HeLa cells and in Rattus norvegicus hepatocytes with localization in the Golgi membranes

704779

transformation of a modified plasmid into Escherichia coli JC201 enzyme-deficient strain and transformation of the plasmid DNA containing the cDNA BAT2 encoding the plastidial enzyme into the Escherichia coli XL-1 red mutator strain

transformation of plants of spring high-erocic acid rape, Hero, with a construct carrying the Limnanthes douglasii enzyme, LAT2, under control of the napin promoter. Study of the correlation between 22:1 accumulation at sn-2, production of trierucin and the expression of the transgene as detected by Western blot

Limnanthes douglasii

487208

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

expression of isoform AGPAT11 mRNA is significantly upregulated in human breast, cervical, and colorectal cancer tissues

Q7L5N7

720175

expression of the enzyme in the testis increases significantly in an age-dependent manner, which is regulated by induction through beta-estradiol, in TM4 cells

isoform LPAAT3 is induced during germ cell maturation. Differentiation of mouse GC-2spd(ts) spermatocytes into spermatides up-regulates isoform LPAAT3 mRNA, increases the amount of polyunsaturated phospholipids, and shifts the specificity for the incorporation of docosahexaenoic acid toward phosphatidylcholine and phosphatidylethanolamine

719440

the transcript level is constant during coconut endosperm development

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

C247Y

loss of the ability of the plastidial enzyme to restore growth to JC201, suggests that C247 is essential for activity

D199A

the mutant fails to grow beyond 0.04 D600 after transfer to the non-permissive temperature. D199 is essential for the ability of the plastidial enzyme to complement the defective acyltransferase activity of JC201

E266A

the plastidial enzyme containing this mutation is unable to complement the temperature-sensitive phenotype of JC201, indicating an essential role for this acidic residue

G140S

loss of the ability of the plastidial enzyme to restore growth to JC201, suggests that G140 is essential for activity

H194A

the mutant fails to grow beyond 0.04 D600 after transfer to the non-permissive temperature. H194 is essential for the ability of the plastidial enzyme to complement the defective acyltransferase activity of JC201

K84Q

greatly impaired growth rate, possibly indicating an importance of the N-terminus of the mature protein in determining enzyme activity

L203P

loss of the ability of the plastidial enzyme to restore growth to JC201, suggests that L203 is essential for activity

P187L

loss of the ability of the plastidial enzyme to restore growth to JC201, suggests that P187 is essential for activity

R92G

greatly impaired growth rate, possibly indicating an importance of the N-terminus of the mature protein in determining enzyme activity

T122A

the change increases the substrate specificity in vitro for oleoyl-CoA and linoleoyl-CoA

487205

T122L

the change increases the substrate specificity in vitro for lignoceroyl-CoA

487205

140delF

34.8% of wild-type activity

221delGT

5% of wild-type activity

252delMRT

3.2% of wild-type activity

A239V

90% of wild-type activity

D168R

site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type AGPAT8

671597

D180fsX251

7.7% of wild-type activity

E260K

site-directed mutagenesis, mutation found in Chanarin-Dorfman syndrome patients, mutation without significant effect on acyltransferase activity

G136R

40.3% of wild-type activity

I208A

mutant of human AGPAT1

L228P

23.7% of wild-type activity

P207L

mutant of human AGPAT1

Q130P

site-directed mutagenesis, mutation found in Chanarin-Dorfman syndrome patients, mutation without significant effect on acyltransferase activity

R149K

mutant of human AGPAT1

R68X

14.2% of wild-type activity

T180S

mutant of human AGPAT1

V167fsX228

14.9% of wild-type activity

additional information

12 entries

Please wait a moment until the data is sorted. This message will disappear when the data is sorted.

487179

by mixing 1 vol. of the fraction containing the enzyme with 4 vol. of sonicated phosphatidylcholine/phosphatidylglycerol or total E. coli lipid vesicles

487180

reassociation of the solubilized enzyme with soybean phospholipids

487198