Multiple isoforms of metallothionein are expressed in the porcine liver.
Isogenes are highly homologous to each other and are often difficult to ascertain, as has been the case with metallothionein, a metal-binding protein rich in cysteines. Conventional separation of metallothionein isoforms relied on ion exchange chromatography of the proteins, or screening for the sequences from gene libraries. In this study, a combination of RT-PCR and partial protein sequencing is used in the identification of metallothionein isogenes expressed in porcine liver. By this approach, we have identified expressed coding sequences which constitute 10 new isogenes. Of the four known groups of metallothioneins (MT), phylogenetic analyses place these pig isogenes in the MT-1 group, except two which are identified as being closely related to MT-2, and none in groups 3 and 4. The isogenes are thus namedpMT-1a to -1g, andpMT-2a and -2b. While each of the isogene sequences is unique, two isogenes,pMT-1e1 andpMT-1e2, share an identical amino acid sequence, differing only in specific codons. Two others,pMT-1b andpMT-1g, have a cysteine substituted by arginine, the first such sequence ever detected in MT.pMT-2a andpMT-2b are closely aligned with the MT-2 group of vertebrates, in spite of the absence of a characteristic acidic amino acid at position 10 or 11, common in other mammalian metallothioneins.
Huang
M C
MC
Department of Life Science, National Tsing Hua University, Hsinchu 30034, ROC Taiwan. huang@jhunix.hcf.jhu.edu
Pan
P K
PK
Zheng
Z F
ZF
Chen
N C
NC
Peng
J Y
JY
Huang
P C
PC
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D008099:Liver / Q000737:chemistry*; D008667:Metalloproteins / Q000737:chemistry*; D008668:Metallothionein / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017421:Sequence Analysis; D012689:Sequence Homology, Nucleic Acid; D013552:Swine
D008667:Metalloproteins; D010446:Peptide Fragments; D008668:Metallothionein