PMID- 3365687
OWN - NLM
STAT- MEDLINE
DCOM- 19880615
LR  - 20131121
IS  - 0008-5472 (Print)
IS  - 0008-5472 (Linking)
VI  - 48
IP  - 11
DP  - 1988 Jun 1
TI  - Characterization of cytosolic aldehyde dehydrogenase from cyclophosphamide 
      resistant L1210 cells.
PG  - 2963-8
AB  - The cytosolic aldehyde dehydrogenase (ALDH) isozyme from cyclophosphamide (CPA) 
      resistant L1210 cells (L1210/CPA) was purified to apparent homogeneity using 
      ternary enzyme complex-dye ligand chromatography. The purified isozyme migrates 
      as a single band at Mr 51,000 in sodium dodecyl sulfate polyacrylamide gel 
      electrophoresis and as a single charge species at isoelectric point = 5.8 in 
      isoelectric focusing. Micromolar Km values were estimated with both 
      propionaldehyde (Km = 5 microM) and 4-hydroxy cyclophosphamide (4-OH CPA) (Km = 4 
      microM) as substrates, indicating that this isozyme is capable of oxidizing the 
      activated cyclophosphamide intermediate 4-hydroxy CPA/aldophosphamide to 
      carboxyphosphamide. This isozyme is also potently inhibited by disulfiram (Ki = 6 
      microM) and 4-(diethylamino)benzaldehyde (Ki = 0.04 microM). Both of these 
      inhibitors are capable of sensitizing L1210/CPA cells to activated CPA in 
      clonogenic survival assays. Thus, the increased levels of only the cytosolic ALDH 
      isoform in L1210/CPA cells appear to be the single phenotypic difference 
      necessary for conferring resistance to CPA. Monospecific antibodies to the 
      L1210/CPA isozyme have been used in Western blot analysis to detect nanogram 
      levels of ALDH in cell and tissue extracts. These antibodies cross-react with the 
      cytosolic isozyme in P388/CPA cells, mouse liver, mouse small intestine, and the 
      1C1C7 hepatoma cell line, whereas no ALDH is detected in sensitive L1210 or P388 
      cells. Also, these antibodies show little cross-reactivity with the mitochondrial 
      isozyme from mouse liver or 1C1C7 cells. From immunological and inhibitor 
      characterization, the soluble ALDH isozyme in L1210/CPA cells appears identical 
      to the normal mouse tissue isozyme.
FAU - Russo, J E
AU  - Russo JE
AD  - Pharmacology Laboratory, Johns Hopkins Oncology Center, Baltimore, Maryland 
      21205.
FAU - Hilton, J
AU  - Hilton J
LA  - eng
GR  - CA-16783/CA/NCI NIH HHS/United States
GR  - CA-36966/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Cancer Res
JT  - Cancer research
JID - 2984705R
RN  - 8N3DW7272P (Cyclophosphamide)
RN  - EC 1.2.1.3 (Aldehyde Dehydrogenase)
SB  - IM
MH  - Aldehyde Dehydrogenase/*isolation & purification/metabolism
MH  - Animals
MH  - Cyclophosphamide/pharmacology
MH  - Cytosol/enzymology
MH  - Drug Resistance
MH  - Kinetics
MH  - Leukemia L1210/*enzymology
MH  - Mice
MH  - Mitochondria/enzymology
MH  - Tumor Stem Cell Assay
EDAT- 1988/06/01 00:00
MHDA- 1988/06/01 00:01
CRDT- 1988/06/01 00:00
PHST- 1988/06/01 00:00 [pubmed]
PHST- 1988/06/01 00:01 [medline]
PHST- 1988/06/01 00:00 [entrez]
PST - ppublish
SO  - Cancer Res. 1988 Jun 1;48(11):2963-8.