Purification and determination of the binding site of lactate dehydrogenase from chicken breast muscle on immobilized ferric ions.
Lactate dehydrogenase from chicken breast muscle was purified to homogeneity in one step by immobilized metal ion affinity chromatography. The purified enzyme was used to localize the binding site to immobilized Fe(III) ions. After cyanogen bromide degradation and digestion with trypsin, small enzyme fragments capable of binding to immobilized Fe(III) ions were obtained. It is proposed that several histidyl groups are involved in the binding.
Chaga
G
G
Biochemical Separation Centre, Uppsala University, Sweden.
Andersson
L
L
Porath
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D005290:Ferric Compounds / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D005290:Ferric Compounds; D010446:Peptide Fragments; D006639:Histidine; D007770:L-Lactate Dehydrogenase; D014357:Trypsin; D003488:Cyanogen Bromide