Cloning, expression and characterisation of a new human low Mr phosphotyrosine protein phosphatase originating by alternative splicing.
RT-PCR experiments on RNA from K562 and HepG2 cells and from human placenta led to the isolation of a novel cDNA, a further alternative splicing product of the primary transcript of low Mr phosphotyrosine phosphatase (LMW-PTP), already known to produce isoforms 1 and 2. This new transcript represents 15-20% of the total LMW-PTP mRNA in the cell. This novel cDNA codifies for a protein that we have named SV3 (splicing variant 3): the deduced protein sequence presents the first 49 residues identical to those of isoform 1, followed by 24 unrelated amino acids, due to a frameshift introduced at the novel exon-exon boundary. The SV3 protein, expressed in E. coli is enzymatically inactive, most probably because unfolded, as suggested by far-UV circular dichroism (CD) experiments. SV3 protein appears to possess the characteristics of an unstructured polypeptide chain lacking the packing of side chain residues and the secondary structure level that are typical of globular proteins. This protein could represent an inactive variant of the human LMW-PTP.
Modesti
A
A
Department of Biochemical Sciences, University of Florence, Italy.
Marzocchini
R
R
Raugei
G
G
Chiti
F
F
Sereni
A
A
Magherini
F
F
Ramponi
G
G
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006801:Humans; D007527:Isoenzymes; D008969:Molecular Sequence Data; D017027:Protein Tyrosine Phosphatases / Q000235:genetics* / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger / Q000378:metabolism
D018076:DNA, Complementary; D007527:Isoenzymes; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; C490429:ACP1 protein, human; D017027:Protein Tyrosine Phosphatases