Profiling the changes in signaling pathways in ascorbic acid/β-glycerophosphate-induced osteoblastic differentiation.
Despite numerous reports on the ability of ascorbic acid and β-glycerophosphate (AA/β-GP) to induce osteoblast differentiation, little is known about the molecular mechanisms involved in this phenomenon. In this work, we used a peptide array containing specific consensus sequences (potential substrates) for protein kinases and traditional biochemical techniques to examine the signaling pathways modulated during AA/β-GP-induced osteoblast differentiation. The kinomic profile obtained after 7 days of treatment with AA/β-GP identified 18 kinase substrates with significantly enhanced or reduced phosphorylation. Peptide substrates for Akt, PI3K, PKC, BCR, ABL, PRKG1, PAK1, PAK2, ERK1, ERBB2, and SYK showed a considerable reduction in phosphorylation, whereas enhanced phosphorylation was observed in substrates for CHKB, CHKA, PKA, FAK, ATM, PKA, and VEGFR-1. These findings confirm the potential usefulness of peptide microarrays for identifying kinases known to be involved in bone development in vivo and in vitro and show that this technique can be used to investigate kinases whose function in osteoblastic differentiation is poorly understood.
Chaves Neto
Antonio Hernandes
AH
Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas, 13083-970 Campinas, SP, Brazil.
Queiroz
Karla Cristiana
KC
Milani
Renato
R
Paredes-Gamero
Edgar Julian
EJ
Justo
Giselle Zenker
GZ
Peppelenbosch
Maikel P
MP
Ferreira
Carmen Veríssima
CV
D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D005994:Glycerophosphates / Q000378:metabolism*; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction
D005994:Glycerophosphates; D001205:Ascorbic Acid; C031463:beta-glycerophosphoric acid