PMID- 6784763
OWN - NLM
STAT- MEDLINE
DCOM- 19810723
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 20
IP  - 7
DP  - 1981 Mar 31
TI  - Argininosuccinate lyase: purification and characterization from human liver.
PG  - 2056-60
AB  - Argininosuccinate lyase has been purified to near homogeneity and partially 
      characterized from extracts of human liver. The purified enzyme had a specific 
      activity of 10.3 mumol min-1 mg-1 in the forward, argininosuccinate cleaving, 
      reaction and 8.0 mumol min-1 mg-1 in the reverse reaction. On the basis of 
      electrophoretic mobility in sodium dodecyl sulfate containing polyacrylamide 
      gels, the protein had a minimum molecular weight of 49 000. Sedimentation 
      equilibrium centrifugation revealed a molecular weight of 187 000. On the basis 
      of these data, the enzyme appears to be a tetramer composed of subunits of 
      identical molecular weight. The Km values were about 0.20 mM for 
      argininosuccinate, 5.3 mM for fumarte, and 3.0 mM for arginine. The enzyme 
      exhibited normal Michaelis-Menten kinetics, and guanosine 5'-triphosphate (GTP) 
      had no affect on the activity of the enzyme. With the exception of its kinetic 
      properties, the enzyme is very similar to the beef liver enzyme. Antibodies were 
      prepared in rabbits and were specific for the human protein, reacting only 
      slightly with the beef liver enzyme and not at all with the rat liver enzyme. The 
      antibodies reacted identically with purified enzyme and enzyme in extracts of 
      human skin fibroblasts. Immunoadsorption of crude human liver extracts followed 
      by analysis of the immunoprecipitates by sodium dodecyl sulfate gel 
      electrophoresis showed only one protein band which corresponded in mobility to 
      purified argininosuccinate lyase, demonstrating that the antibodies were specific 
      for argininosuccinate lyase.
FAU - O'Brien, W E
AU  - O'Brien WE
FAU - Barr, R H
AU  - Barr RH
LA  - eng
GR  - AM 25938/AM/NIADDK NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Macromolecular Substances)
RN  - EC 4.- (Lyases)
RN  - EC 4.3.2.1 (Argininosuccinate Lyase)
SB  - IM
MH  - Animals
MH  - Argininosuccinate Lyase/*isolation & purification/metabolism
MH  - Cattle
MH  - Fibroblasts/enzymology
MH  - Humans
MH  - Immunoassay
MH  - Immunodiffusion
MH  - Kinetics
MH  - Liver/*enzymology
MH  - Lyases/*isolation & purification
MH  - Macromolecular Substances
MH  - Molecular Weight
MH  - Skin/enzymology
EDAT- 1981/03/31 00:00
MHDA- 1981/03/31 00:01
CRDT- 1981/03/31 00:00
PHST- 1981/03/31 00:00 [pubmed]
PHST- 1981/03/31 00:01 [medline]
PHST- 1981/03/31 00:00 [entrez]
AID - 10.1021/bi00510a049 [doi]
PST - ppublish
SO  - Biochemistry. 1981 Mar 31;20(7):2056-60. doi: 10.1021/bi00510a049.