Characterization of metallothioneins (MT-I and MT-II) in the yak.

The cDNA-encoding sequences for yak metallothionein isoforms I (MT-I) and II (MT-II) were amplified and cloned by reverse-transcription PCR to characterize the nucleotide sequence and protein structure of metallothionein in the yak. The cDNA sequences of MT-I and MT-II were subjected to BLAST searching at the National Center for Biotechnology Information, and the results indicated that the nucleotide sequences of yak MT-I and MT-II, when compared among different species of mammals, are highly conserved. The yak open reading frames have a length of 183 nucleotides, which encode for yak MT-I and MT-II proteins of 61 AA, respectively. Analysis of hydrophobicity, trans-membrane region, and signal peptides suggested that metallothioneins of the yak are nonsecretory proteins. There were several conserved tripeptide sequences, such as C-X-C, C-C-X-C-C, and C-X-X-C (X designates AA excluding cysteine in MT-I and MT-II), and they are highly conserved in their evolution. By homologous comparative modeling, we predicted the molecular spatial structures of yak MT-I and MT-II, which are composed of alpha- and beta-domains that are linked by the conserved tripeptide Lys(30)-Lys(31)-Ser(32) (KKS).