L-(+)-Cysteine → (2R)-2-Acetamido-3-({(2R,3S,4R)-3-hydroxy-2-[(1S)-1-hydroxy-2-methylpropyl]-4-methyl-5-oxopyrrolidine-2-carbonyl}sulfanyl)propanoic acidLactacystin

HRID 1292600Notes

Reaction equation

HRID 1292600 reaction equation
CC(C[C@H](N[C:9]([C@@H:11]([NH:16][C:17]([C@@H:19](NC(OCC1C=CC=CC=1)=O)[CH2:20]C(C)C)=[O:18])[CH2:12][CH:13]([CH3:15])[CH3:14])=[O:10])C=O)C.N[C@H:36]([C:41]([OH:43])=O)CCSC.[NH2:44][C@H:45]([C:48]([OH:50])=[O:49])[CH2:46][SH:47].CCCCCCCCCCC[CH2:62][O:63]S([O-])(=O)=O.[Na+].C(O)C(N)(CO)C[OH:72]>CS(C)=O>[CH3:20][C@H:19]1[C:17](=[O:18])[NH:16][C@:11]([C:62]([S:47][CH2:46][C@H:45]([NH:44][C:41]([CH3:36])=[O:43])[C:48]([OH:50])=[O:49])=[O:63])([C@@H:12]([OH:72])[CH:13]([CH3:14])[CH3:15])[C@H:9]1[OH:10]

Participants

6

Reactant 4

HCID 5862L-(+)-CysteineHCID 6137L-MethionineHCID 462382MG132HCID 88195029HCID 88195029, compound

Product 1

HCID 6610292(2R)-2-Acetamido-3-({(2R,3S,4R)-3-hydroxy-2-[(1S)-1-hydroxy-2-methylpropyl]-4-methyl-5-oxopyrrolidine-2-carbonyl}sulfanyl)propanoic acidLactacystin

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent

Procedure

COS cells were plated onto 100 mm dishes, then, with DEAE-Destran, either mock transfected, or transfected with 3 μg of pcDNA plasmid containing the human p105 cDNA. Forty-eight hrs after transfection, cells were pretreated for 1 hour with 0.5% DMSO, 50 μM calpain inhibitor II, 50 μM MG132, or 10 μM β-lactone. Cells were then pulse labelled with 250 uCi/plate of 35S-methionine/cysteine for 15 minutes, and either harvested immediately or followed by a 2 hour chase with excess unlabelled methionine and cysteine. Cells were then lysed with SDS/Tris, and proteins were immunoprecipitated with anti-p50 antibodies (recognize the N-terminal half of the p105 protein). These proteins were then separated on a 10% SDS-polyacrylamide gel, which was then fixed, dried, and exposed to autoradiographic film.

Workup

  1. additioncontaining the human p105 cDNA
  2. customThese proteins were then separated on a 10% SDS-polyacrylamide gel, which
  3. customdried

Sources & evidence

ORD record
ord-f5f88b14dca64cb0bc087d9ca62d2594
Source dataset
uspto-grants-1998_05
Patent number
US05756764

Reaction notes

Loading…