Stratifin, a keratinocyte specific 14-3-3 protein, harbors a pleckstrin homology (PH) domain and enhances protein kinase C activity.
The intrinsic signal(s) responsible for the onset of human keratinocyte terminal differentiation is not yet fully understood. Evidence has been recently accumulated linking the phospholipase-mediated activation of protein kinase C to the coordinate changes in gene expression occurring during keratinocyte terminal differentiation. Here we report the purification of a keratinocyte-derived protein enhancing protein kinase C enzymatic activity. The stimulator eluted as a peak with estimated molecular mass of approximately 70 kDa, while analysis by SDS-PAGE showed a 30 kDa protein migrating as a distinct doublet, suggesting the formation of a 30 kDa homodimer. The amino acid sequence analysis allowed the unambigous identification of the protein kinase C stimulator as a mixture of the highly homologous sigma (stratifin) and zeta isoforms of 14-3-3 proteins, which are homodimers of identical 30 kDa subunits. Mono Q anion exchange chromatography and immunoblot analysis further confirmed that stratifin enhances protein kinase C activity. Stratifin was originally sequenced from a human keratinocyte protein database, but its function was unknown. The pleckstrin homology domain has been recently related to protein translocation to the cell membrane as well as to functional interactions of intracellular proteins involved in signal transduction. We show here that stratifin (and 14-3-3 zeta) harbors a pleckstrin homology domain, and the consequent functional implications will be discussed.
Dellambra
E
E
National Cancer Institute/Advanced Biotechnology Center, Genova, Italy.
Patrone
M
M
Sparatore
B
B
Negri
A
A
Ceciliani
F
F
Bondanza
S
S
Molina
F
F
Cancedda
F D
FD
De Luca
M
M
D048948:14-3-3 Proteins; D016475:3T3 Cells / Q000201:enzymology; D000595:Amino Acid Sequence; D000818:Animals; D000837:Anion Exchange Resins; D014408:Biomarkers, Tumor; D001798:Blood Proteins / Q000648:ultrastructure*; D001921:Brain / Q000201:enzymology; D002454:Cell Differentiation / Q000502:physiology; D002852:Chromatography, Ion Exchange; D004789:Enzyme Activation; D000078404:Epidermal Cells; D005092:Exonucleases; D005095:Exoribonucleases; D006801:Humans; D015151:Immunoblotting; D015603:Keratinocytes / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D011493:Protein Kinase C / Q000378:metabolism*; D011506:Proteins / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats; D012117:Resins, Synthetic; D015398:Signal Transduction / Q000502:physiology
D048948:14-3-3 Proteins; D000837:Anion Exchange Resins; D014408:Biomarkers, Tumor; D001798:Blood Proteins; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D011506:Proteins; D012117:Resins, Synthetic; C039077:platelet protein P47; C042855:Mono Q; D011493:Protein Kinase C; D005092:Exonucleases; D005095:Exoribonucleases; C413442:SFN protein, human