Identification, synthesis and properties of a consensus peptide recognized by a monoclonal antibody directed to various type I cytokine receptors.
Previous works demonstrated that the monoclonal antibody (MAb) called R7B4 is directed to an epitope shared by receptors for lactogenic and somatogenic hormones as well as interleukins 2 and 6 (IL-2 and IL-6). The MAb inhibited the biological effects of those hormones and cytokines by impairing their binding to receptors. It is known that the receptors for growth hormones (GH), prolactins (PRL), IL-2, and IL-6 pertain to the type I cytokine receptor family, sharing the common motif WSXWS or the homologous F(Y)GEFS. Thus, a set of 34 decapeptides corresponding to diverse receptors containing those sequences were synthesized by the PEPSCAN method and their reactions with MAb R7B4 were measured by ELISA. The MAb significantly recognized 21 peptides, allowing us to establish the consensus sequence HGYWSEWSPE as a portion of the R7B4 epitope. The consensus peptide was synthesized and purified by conventional methods, and its capacity to bind to MAb R7B4 paratope confirmed. Moreover, polyclonal Ab to the peptide elicited in mice were able to inhibit the hGH binding to lactogenic, somatogenic and human specific liver receptors. This fact suggests that the consensus peptide could be used as an immunogen to produce anti-hGH receptor Ab behaving as hormone or cytokine antagonists in certain pathological conditions.
Belloc
Carlos G
CG
Instituto de Química y Fisicoquímica Biológicas, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Longhi
Silvia A
SA
Peña
Clara
C
Blank
Viviana A
VA
Miranda
Marta E
ME
Retegui
Lilia A
LA
D000595:Amino Acid Sequence; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D030541:Databases, Genetic; D004797:Enzyme-Linked Immunosorbent Assay; D013006:Growth Hormone / Q000378:metabolism; D006801:Humans; D007114:Immunization; D007457:Iodine Radioisotopes; D051379:Mice; D008807:Mice, Inbred BALB C; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D011817:Rabbits; D051381:Rats; D018121:Receptors, Cytokine / Q000037:antagonists & inhibitors*
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D009479:Neuropeptides; D018121:Receptors, Cytokine; D013006:Growth Hormone