The proline-rich focal adhesion and microfilament protein VASP is a ligand for profilins.
Profilins are small proteins that form complexes with G-actin and phosphoinositides and are therefore considered to link the microfilament system to signal transduction pathways. In addition, they bind to poly-L-proline, but the biological significance of this interaction is not yet known. The recent molecular cloning of the vasodilator-stimulated phosphoprotein (VASP), an established in vivo substrate of cAMP- and cGMP-dependent protein kinases, revealed the presence of a proline-rich domain which prompted us to investigate a possible interaction with profilins. VASP is a microfilament and focal adhesion associated protein which is also concentrated in highly dynamic regions of the cell cortex. Here, we demonstrate that VASP is a natural proline-rich profilin ligand. Human platelet VASP bound directly to purified profilins from human platelets, calf thymus and birch pollen. Moreover, VASP and a novel protein were specifically extracted from total cell lysates by profilin affinity chromatography and subsequently eluted either with poly-L-proline or a peptide corresponding to a proline-rich VASP motif. Finally, the subcellular distributions of VASP and profilin suggest that both proteins also interact within living cells. Our data support the hypothesis that profilin and VASP act in concert to convey signal transduction to actin filament formation.
Reinhard
M
M
Medizinische Universitätsklinik, Klinische Biochemie und Pathobiochemie, Würzburg, Germany.
Giehl
K
K
Abel
K
K
Haffner
C
C
Jarchau
T
T
Hoppe
V
V
Jockusch
B M
BM
Walter
U
U
D000595:Amino Acid Sequence; D000818:Animals; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002451:Cell Compartmentation; D002465:Cell Movement; D002846:Chromatography, Affinity; D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011058:Pollen / Q000737:chemistry; D051304:Profilins; D011485:Protein Binding; D051381:Rats; D017421:Sequence Analysis; D015398:Signal Transduction; D012867:Skin / Q000166:cytology / Q000378:metabolism; D013045:Species Specificity; D000098662:Vasodilator-Stimulated Phosphoprotein; D000099149:Ezrin
D015815:Cell Adhesion Molecules; D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D051304:Profilins; D000098662:Vasodilator-Stimulated Phosphoprotein; D000099149:Ezrin; C495740:Pfn1 protein, rat