PMID- 8204587
OWN - NLM
STAT- MEDLINE
DCOM- 19940708
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 33
IP  - 21
DP  - 1994 May 31
TI  - Inhibition of glyceraldehyde-3-phosphate dehydrogenase by pentalenolactone. 2. 
      Identification of the site of alkylation by tetrahydropentalenolactone.
PG  - 6524-30
AB  - Incubation of rabbit muscle glyceraldehyde-3-phosphate dehydrogenase (GAPDH) with 
      the antibiotic pentalenolactone (3) results in time-dependent, irreversible 
      inhibition of GAPDH by modification of a single Cys residue in each subunit of 
      the homotetrameric enzyme. Reduction of pentalenolactone with tritium gas gave 
      [2,3,7,8-3H4]tetrahydropentalenolactone (7), which also exhibited time-dependent, 
      irreversible inactivation of GAPDH. The site of covalent attachment of 7 was 
      determined. Tryptic digestion of inactivated GAPDH and purification of the 
      resultant products by reverse-phase HPLC gave a single labeled peptide. Amino 
      acid sequence analysis of the radioactive peptide gave 
      Ile-Val-Ser-Asn-Ala-Ser-X-Thr-Thr-Asn-(...). This sequence is identical to the 
      highly conserved region from Ile-143 to Asn-152 in pig muscle GAPDH, except for 
      the active site Cys-149 to which the tetrahydropentalenolactone was covalently 
      bound. Molecular modeling was used to compare both pentalenolactone (3) and 
      heptelidic acid (4), a mechanistically related inactivator of GAPDH, with the 
      normal substrate, glyceraldehyde 3-phosphate (1). Finally, pentalenolactone was 
      shown by reaction with model thiols to undergo epoxide ring opening exclusively 
      by nucleophilic attack at the primary carbon, C-10.
FAU - Cane, D E
AU  - Cane DE
AD  - Department of Chemistry, Brown University, Providence, Rhode Island 02912.
FAU - Sohng, J K
AU  - Sohng JK
LA  - eng
GR  - GM-22172/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Sesquiterpenes)
RN  - 0 (Sulfhydryl Compounds)
RN  - 156900-92-4 (tetrahydropentalenolactone)
RN  - 31501-48-1 (arenaemycin E)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
SB  - IM
MH  - Alkylation
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - Chromatography, High Pressure Liquid
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/*antagonists & inhibitors
MH  - Magnetic Resonance Spectroscopy
MH  - Molecular Sequence Data
MH  - Muscles/enzymology
MH  - Rabbits
MH  - Sesquiterpenes/chemistry/*pharmacology
MH  - Sulfhydryl Compounds/chemistry
EDAT- 1994/05/31 00:00
MHDA- 1994/05/31 00:01
CRDT- 1994/05/31 00:00
PHST- 1994/05/31 00:00 [pubmed]
PHST- 1994/05/31 00:01 [medline]
PHST- 1994/05/31 00:00 [entrez]
AID - 10.1021/bi00187a020 [doi]
PST - ppublish
SO  - Biochemistry. 1994 May 31;33(21):6524-30. doi: 10.1021/bi00187a020.