Isolation, characterization, and determination of human liver (copper/zinc) metallothionein.
A copper-containing protein was purified from the liver of a patient with primary biliary cirrhosis by a combination of gel filtration and anion exchange chromatography. This copper-protein had UV absorption and emission spectra, an amino acid composition, and a molecular mass which were characteristic for metallothionein (MT). From 8 livers (3 control, 1 fetal and 4 primary biliary cirrhosis) MT was extracted with non-reducing buffer and centrifuged, and pellets were re-extracted with a 1% 2-mercaptoethanol-containing buffer. The non-reducing buffer extracted a predominantly copper-containing MT from the livers of patients with primary biliary cirrhosis and a predominantly zinc-containing MT from control livers and the fetal liver. Only from the fetal liver was a copper/zinc containing MT solubilized during the re-extraction with 2-mercaptoethanol-containing buffer. These results indicate that human MT is a unique metalloprotein with age and disease-dependent characteristics.
Mulder
T P
TP
Department Gastroenterology and Hepatology, University Hospital, Leiden, The Netherlands.
Janssens
A R
AR
Verspaget
H W
HW
Lamers
C B
CB
D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D003300:Copper; D006801:Humans; D008099:Liver / Q000032:analysis*; D008105:Liver Cirrhosis, Biliary / Q000378:metabolism; D008668:Metallothionein / Q000302:isolation & purification*; D008970:Molecular Weight; D015032:Zinc
D000596:Amino Acids; D003300:Copper; D008668:Metallothionein; D015032:Zinc