Signalling properties of an HIV-encoded angiogenic peptide mimicking vascular endothelial growth factor activity.
HIV-1 expresses a multifunctional protein called TAT (trans-acting transcriptional activator), the function of which in vivo is tightly correlated with the incidence of Kaposi's sarcoma in AIDS patients. TAT is angiogenic and apparently binds to receptors specific for vascular endothelial growth factor (VEGF). Amino acids 46-60 of HIV-TAT, known as the basic peptide, have been shown to be responsible for its functional interaction with VEGF receptors. To characterize further the binding properties of this peptide, its coding sequence was fused to the reading frame of bacterial thioredoxin, allowing the production of large amounts of chimaeric polypeptides in bacteria in a biologically active form. Binding of chimaeric proteins to VEGF receptors was studied in vitro in endothelial cell cultures expressing either of the two receptors. Chimaeric thioredoxin proteins carrying the basic domain of TAT bound to both VEGF receptors with affinities similar to those of HIV-TAT or VEGF. Interestingly, these polypeptides competed only partially with VEGF for receptor binding, implying different binding sites for the TAT peptide and VEGF. This suggests that TAT binds VEGF receptors at new sites that might be useful targets for pharmacological intervention during pathological angiogenesis. The thioredoxin/basic-peptide chimaeras are functional agonists that mediate VEGF receptor signalling: (1) they stimulate the growth of endothelial cells; (2) together with basic fibroblast growth factor they cause tube formation of endothelial cells in collagen gels; (3) they induce blood vessel formation on the chicken chorioallantoic membrane; and (4) they activate VEGF receptor kinase and mitogen-activated protein kinase activity.
Scheidegger
P
P
Institute of Medical Radiobiology of the University of Zürich and the Paul Scherrer Institute, CH-5232 Villigen-PSI, Switzerland.
Weiglhofer
W
W
Suarez
S
S
Console
S
S
Waltenberger
J
J
Pepper
M S
MS
Jaussi
R
R
Ballmer-Hofer
K
K
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D017931:DNA Primers; D004285:Dogs; D016228:Endothelial Growth Factors / Q000235:genetics / Q000502:physiology*; D015696:Gene Products, tat / Q000502:physiology*; D006678:HIV / Q000235:genetics*; D006801:Humans; D008222:Lymphokines / Q000235:genetics / Q000502:physiology*; D018716:Molecular Mimicry; D018919:Neovascularization, Physiologic; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D054322:tat Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D016228:Endothelial Growth Factors; D015696:Gene Products, tat; D008222:Lymphokines; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D054322:tat Gene Products, Human Immunodeficiency Virus