Phosphoproteome reveals an atlas of protein signaling networks during osteoblast adhesion.
Cell adhesion on surfaces is a fundamental process in the emerging biomaterials field and developmental events as well. However, the mechanisms regulating this biological process in osteoblasts are not fully understood. Reversible phosphorylation catalyzed by kinases is probably the most important regulatory mechanism in eukaryotes. Therefore, the goal of this study is to assess osteoblast adhesion through a molecular prism under a peptide array technology, revealing essential signaling proteins governing adhesion-related events. First, we showed that there are main morphological changes on osteoblast shape during adhesion up to 3 h. Second, besides classical proteins activated upon integrin activation, our results showed a novel network involving signaling proteins such as Rap1A, PKA, PKC, and GSK3beta during osteoblast adhesion on polystyrene. Third, these proteins were grouped in different signaling cascades including focal adhesion establishment, cytoskeleton rearrangement, and cell-cycle arrest. We have thus provided evidence that a global phosphorylation screening is able to yield a systems-oriented look at osteoblast adhesion, providing new insights for understanding of bone formation and improvement of cell-substratum interactions. Altogether, these statements are necessary means for further intervention and development of new approaches for the progress of tissue engineering.
Milani
Renato
R
Department of Biochemistry, Institute of Biology, University of Campinas (UNICAMP), 13083-970 Campinas, São Paulo, Brazil.
Ferreira
Carmen V
CV
Granjeiro
José M
JM
Paredes-Gamero
Edgar J
EJ
Silva
Rodrigo A
RA
Justo
Giselle Z
GZ
Nader
Helena B
HB
Galembeck
Eduardo
E
Peppelenbosch
Maikel P
MP
Aoyama
Hiroshi
H
Zambuzzi
Willian F
WF
D051339:Actin Depolymerizing Factors / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D020816:Amino Acid Motifs; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape; D003599:Cytoskeleton / Q000378:metabolism; D022001:Focal Adhesions / Q000201:enzymology; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism; D040081:Protein Array Analysis; D020543:Proteome / Q000378:metabolism*; D015203:Reproducibility of Results; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors
D051339:Actin Depolymerizing Factors; D000199:Actins; D010750:Phosphoproteins; D020543:Proteome; D012694:Serine; D010770:Phosphotransferases