5'-Atp → Tetradecanoyl-coa

HRID 373594

Reaction equation

[C:1]([O-:16])(=O)[CH2:2][CH2:3][CH2:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH2:10][CH2:11][CH2:12][CH2:13][CH3:14].P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.[C@@H:48]1([N:86]2[C:95]3[N:94]=[CH:93][N:92]=[C:90]([NH2:91])[C:89]=3[N:88]=[CH:87]2)[O:85][C@H:57]([CH2:58][O:59][P:60]([O:63][P:64]([O:67][CH2:68][C:69]([C@H:72]([C:74]([NH:76][CH2:77][CH2:78][C:79]([NH:81][CH2:82][CH2:83][SH:84])=[O:80])=[O:75])[OH:73])([CH3:71])[CH3:70])([OH:66])=[O:65])([OH:62])=[O:61])[C@@H:51]([O:52][P:53]([OH:56])([OH:55])=[O:54])[C@H:49]1[OH:50]>>[C:1]([S:84][CH2:83][CH2:82][NH:81][C:79](=[O:80])[CH2:78][CH2:77][NH:76][C:74](=[O:75])[C@H:72]([OH:73])[C:69]([CH3:70])([CH3:71])[CH2:68][O:67][P:64]([OH:66])(=[O:65])[O:63][P:60]([OH:62])(=[O:61])[O:59][CH2:58][C@H:57]1[O:85][C@@H:48]([N:86]2[C:95]3[N:94]=[CH:93][N:92]=[C:90]([NH2:91])[C:89]=3[N:88]=[CH:87]2)[C@H:49]([OH:50])[C@@H:51]1[O:52][P:53]([OH:56])([OH:55])=[O:54])(=[O:16])[CH2:2][CH2:3][CH2:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH2:10][CH2:11][CH2:12][CH2:13][CH3:14]

Participants

Reactant 3

HCID 59575'-Atp

HCID 87642Coenzyme A

HCID 4075158Myristate

Product 1

HCID 11966124Tetradecanoyl-coa

Procedure

The compounds are then tested on the supernatant in competition with the peptide substrate Gly-Asn-(Ala)4 -(Arg)2 ("G8R") according to the conditions which follow, as described by TOWLER et al. (PNAS, (1987), 84, 2708-2711). Myristoylcoenzyme A is synthesized enzymatically immediately before use by incubation of myristate with ATP, Coenzyme A (lithium salt) and Pseudomonas Acyl-Coa synthetase (20 min at 30° C.). The peptide substrate and the biological source as obtained above are then added. The mixture is incubated for 10 min at 37° C.; the reaction is then stopped with 110 μl of methanol and 10 μl of trichloroacetic acid. The medium is left at 4° C. for 10 min and then centrifuged at 10,000 g for 5 min. An aliquot (30 μl) of the final supernatant is injected into an HPLC system equipped with a μBondapak column (Waters) and developed with a linear gradient as described by Towler et al. (PNAS, (1987), 84, 2708-11). Detection of the acylated peptide is performed using a radioactive line detector (Berthold) by means of the addition of 5 ml/min of scintillation fluid (Zinsser). The acylated peptide is eluted at 85% acetonitrile-TFA/15% H20-TFA.

Sources & evidence

ORD record: ord-f0e1c78075034113a33522964a30baaa
Source dataset: uspto-grants-1992_11
DOI: 10.6084/m9.figshare.5104873.v1
Patent number: US05164414

Linked notes

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