PMID- 11281273
OWN - NLM
STAT- MEDLINE
DCOM- 20010426
LR  - 20190915
IS  - 1096-4959 (Print)
IS  - 1096-4959 (Linking)
VI  - 127
IP  - 4
DP  - 2000 Dec
TI  - Partial purification and characterization of 12-lipoxygenase in bullfrog 
      erythrocytes.
PG  - 563-73
AB  - 12-Lipoxygenase (12-LO) in bullfrog (Rana catesbeiana) erythrocytes was purified 
      partially by ion exchange chromatography and affinity chromatography. Bullfrog 
      12-LO was a single chain protein with a pI of 7.1-7.8 and MW of 7.77 kDa. This 
      enzyme did not show typical Michaelis Menten type kinetics. At low substrate 
      concentrations, it had a lag phase and at higher substrate concentrations, the 
      activity was inhibited. The product of linoleic acid (LA), 13-hydroperoxy-9, 
      11-octadecadienoic acid (13-HpODE), was an activator for the enzyme. When 
      arachidonic acid (AA) was used as substrate, 13-HpODE also affected the Km of 
      bullfrog 12-LO towards AA. The affinity of LA towards bullfrog 12-LO was higher 
      than the affinity of AA. Suicide inactivation was much more rapid than that of 
      any mammalian 12-LO reported. Hemoglobin (Hb) inhibited the activity of 12-LO 
      partially and removing Hb eliminated this inhibition. Both Hb and Met-Hb 
      inhibited the 12-LO activity but did not denatured completely the Hb, suggesting 
      that the inhibition was a direct interaction between 12-LO and Hb protein chain 
      and was not due to competition between 12-LO and Hb for oxygen. This study 
      characterizes bullfrog 12-LO with respect to stability, optimal pH, suicide 
      inactivation and interaction with Hb and provides important evolutionary 
      information about this enzyme.
FAU - Shen, K
AU  - Shen K
AD  - Department of Biology, New Mexico State University, Las Cruces 88003, USA. 
      keqiang.shen@dupontpharma.com
FAU - Herman, C A
AU  - Herman CA
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - England
TA  - Comp Biochem Physiol B Biochem Mol Biol
JT  - Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology
JID - 9516061
RN  - 0 (Hemoglobins)
RN  - 0 (Linoleic Acids)
RN  - 0 (Lipid Peroxides)
RN  - 23017-93-8 (13-hydroperoxy-9,11-octadecadienoic acid)
RN  - 27YG812J1I (Arachidonic Acid)
RN  - EC 1.13.11.31 (Arachidonate 12-Lipoxygenase)
SB  - IM
MH  - Animals
MH  - Arachidonate 12-Lipoxygenase/*analysis/*isolation & purification
MH  - Arachidonic Acid/*metabolism
MH  - Chromatography, Affinity
MH  - Chromatography, High Pressure Liquid
MH  - Chromatography, Ion Exchange
MH  - Chromatography, Thin Layer
MH  - Erythrocytes/*enzymology
MH  - Hemoglobins/metabolism
MH  - Hydrogen-Ion Concentration
MH  - Isoelectric Focusing
MH  - Linoleic Acids/metabolism
MH  - Lipid Peroxides/metabolism
MH  - Molecular Weight
MH  - Rana catesbeiana/*blood
MH  - Subcellular Fractions/metabolism
MH  - Substrate Specificity
MH  - Temperature
EDAT- 2001/04/03 10:00
MHDA- 2001/05/01 10:01
CRDT- 2001/04/03 10:00
PHST- 2001/04/03 10:00 [pubmed]
PHST- 2001/05/01 10:01 [medline]
PHST- 2001/04/03 10:00 [entrez]
AID - 10.1016/s0305-0491(00)00288-1 [doi]
PST - ppublish
SO  - Comp Biochem Physiol B Biochem Mol Biol. 2000 Dec;127(4):563-73. doi: 
      10.1016/s0305-0491(00)00288-1.