The cloning and expression of a human creatine transporter.
A human creatine transporter (hCRT-BS2M) cDNA clone was isolated from a human brainstem/spinal cord using a PCR and phage plaque hybridization based technique. This clone included an open reading frame of 1,905 base pairs(bp) within a 2,283bp cDNA. Northern blot hybridization detected the expression of corresponding mRNAs most prominently in the skeletal muscle, heart and kidney. Peptide sequence analysis of the hCRT-BS2M protein product revealed 12 putative transmembrane domains. The predicted protein sequence further demonstrates that the hCRT-BS2M has highly conserved amino acid identity with the other members of the sodium dependent plasma membrane transporter family. Transient expression of the hCRT-BS2M in COS-7 cells demonstrates sodium dependent [14C]creatine uptake with a KM value of 14.9 +/- 3.0 microM (n = 5) that is attenuated by creatine and selective structural analogues of creatine.
Sora
I
I
Department of Pharmacology, College of Medicine, University of Arizona, Tucson 85724.
Richman
J
J
Santoro
G
G
Wei
H
H
Wang
Y
Y
Vanderah
T
T
Horvath
R
R
Nguyen
M
M
Waite
S
S
Roeske
W R
WR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001933:Brain Stem / Q000378:metabolism*; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003401:Creatine / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007668:Kidney; D007700:Kinetics; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D016133:Polymerase Chain Reaction; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D013116:Spinal Cord / Q000378:metabolism*; D014162:Transfection
D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D026901:Membrane Transport Proteins; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; C080679:creatine transporter; D003401:Creatine