| Autor: |
Y, Li, A P, Valaitis, S P, Latshaw, D, Kwiatkowska, R L, Tripathi, M C, Campbell, R G, Kemp |
| Rok vydání: |
1994 |
| Předmět: |
|
| Zdroj: |
The Journal of biological chemistry. 269(8) |
| ISSN: |
0021-9258 |
| Popis: |
Rabbit brain contains three phosphofructo-1-kinase (PFK) isozymic subunits designated A, B, and C. The primary structures of the first of these two isozyme types have been determined previously. The isozyme C of rabbit brain was isolated by immunoaffinity chromatography and subjected to proteolytic and chemical digestion. A large number of peptides were sequenced, the total number of amino acids identified being equal to about 80% of the total structure. The sequence of the cDNA derived from brain mRNA for C isozyme was determined from polymerase chain reaction fragments synthesized using oligonucleotides designed on the basis of the peptide sequences. The deduced size of the C isozyme was 86,371 Da, slightly larger than PFKs described previously. The amino acid sequence identity with the rabbit A isozyme was 68.9% and a range of identity to other sequenced mammalian PFKs was 67-69%. Using these data plus previously published data on chemical modification, assignments of the 6 organic ligand binding sites of PFK were inferred. The full-length cDNA was cloned into and expressed in Escherichia coli. Phosphofructokinase C was purified to homogeneity from the bacterial extracts. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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