TY - JOUR
T1 - Cloning and nucleotide sequence of a cDNA encoding the rat triosephosphate isomerase
AU - Lee, Kyunglim
AU - Ryu, Jiwon
PY - 1996/12
Y1 - 1996/12
N2 - A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1,348-bp cDNA clone contains 24 bp 5′ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, 547 bp 3′ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI cDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu130, Tyr209, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.
AB - A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1,348-bp cDNA clone contains 24 bp 5′ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, 547 bp 3′ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI cDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu130, Tyr209, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.
KW - cDNA library
KW - Rat skeletal muscle
KW - RT-PCR
KW - TPI
UR - http://www.scopus.com/inward/record.url?scp=20544458108&partnerID=8YFLogxK
U2 - 10.1007/BF02986018
DO - 10.1007/BF02986018
M3 - Article
AN - SCOPUS:20544458108
SN - 0253-6269
VL - 19
SP - 497
EP - 501
JO - Archives of Pharmacal Research
JF - Archives of Pharmacal Research
IS - 6
ER -