PMID- 10212249
OWN - NLM
STAT- MEDLINE
DCOM- 19990603
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 18
DP  - 1999 Apr 30
TI  - Kinetics and inhibition of recombinant human cystathionine gamma-lyase. Toward
      the rational control of transsulfuration.
PG  - 12675-84
AB  - The gene encoding human cystathionine gamma-lyase was cloned from total cellular 
      Hep G2 RNA. Fusion to a T7 promoter allowed expression in Escherichia coli,
      representing the first mammalian cystathionine gamma-lyase overproduced in a
      bacterial system. About 90% of the heterologous gene product was insoluble, and
      renaturation experiments from purified inclusion bodies met with limited success.
      About 5 mg/liter culture of human cystathionine gamma-lyase could also be
      extracted from the soluble lysis fraction, employing a three-step native
      procedure. While the enzyme showed high gamma-lyase activity toward
      L-cystathionine (Km = 0.5 mM, Vmax = 2.5 units/mg) with an optimum pH of 8.2, no 
      residual cystathionine beta-lyase behavior and only marginal reactivity toward
      L-cystine and L-cysteine were detected. Inhibition studies were performed with
      the mechanism-based inactivators propargylglycine, trifluoroalanine, and
      aminoethoxyvinylglycine. Propargylglycine inactivated human cystathionine
      gamma-lyase much more strongly than trifluoroalanine, in agreement with the
      enzyme's preference for C-gamma-S bonds. Aminoethoxyvinylglycine showed slow and 
      tight binding characteristics with a Ki of 10.5 microM, comparable with its
      effect on cystathionine beta-lyase. The results have important implications for
      the design of specific inhibitors for transsulfuration components.
FAU - Steegborn, C
AU  - Steegborn C
AD  - Max-Planck-Institut fur Biochemie, Abteilung Strukturforschung, Am Klopferspitz
      18a, 82152 Planegg-Martinsried, Germany.
FAU - Clausen, T
AU  - Clausen T
FAU - Sondermann, P
AU  - Sondermann P
FAU - Jacob, U
AU  - Jacob U
FAU - Worbs, M
AU  - Worbs M
FAU - Marinkovic, S
AU  - Marinkovic S
FAU - Huber, R
AU  - Huber R
FAU - Wahl, M C
AU  - Wahl MC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA Primers)
RN  - 0 (Recombinant Proteins)
RN  - 70FD1KFU70 (Sulfur)
RN  - EC 4.4.1.1 (Cystathionine gamma-Lyase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Cloning, Molecular
MH  - Cystathionine gamma-Lyase/*antagonists & inhibitors/chemistry/*metabolism
MH  - DNA Primers
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Rats
MH  - Recombinant Proteins/antagonists & inhibitors/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Sulfur/*metabolism
EDAT- 1999/04/23 00:00
MHDA- 1999/04/23 00:01
CRDT- 1999/04/23 00:00
PHST- 1999/04/23 00:00 [pubmed]
PHST- 1999/04/23 00:01 [medline]
PHST- 1999/04/23 00:00 [entrez]
AID - 10.1074/jbc.274.18.12675 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Apr 30;274(18):12675-84. doi: 10.1074/jbc.274.18.12675.