Secondary literature sources for IlGF
The following references were automatically generated.
- Klonisch T, Froehlich C, Tetens F, Fischer B, Hombach-Klonisch S
- Molecular remodeling of members of the relaxin family during primate evolution.
- Mol Biol Evol. 2001; 18: 393-403
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Employing comparative analysis of the cDNA-coding sequences of the unique preprorelaxin of the Afro-lorisiform Galago crassicaudatus and the Malagasy lemur Varecia variegata and the relaxin-like factor (RLF) of G. crassicaudatus, we demonstrated distinct differences in the dynamics of molecular remodeling of both hormones during primate evolution. The lorisiform and lemuriform preprorelaxin sequences encoded identical hormones, providing the first endocrinological evidence for the monophyletic origin of all Strepsirrhini. Structural analysis revealed the lemuriform members of the relaxin family to be potentially bioactive single-gene products. In contrast to the "two-prong" relaxin receptor-binding motif (RELVR) present within the B-domains of other primate relaxins, strepsirrhine relaxin contained a unique "three-prong" motif (RRLIR) with highest sequence homology to the receptor-binding motif of the evolutionarily much older skate relaxin. In contrast to relaxin, the RLF molecule was highly conserved during primate evolution and contained within its B-domain the putative relaxin receptor-binding motif and a pentameric sequence implicated in binding to specific RLF receptors. Mutually exclusive expression of strepsirrhine preprorelaxin and RLF were observed in the fetal villous trophoblast cells of the strepsirrhine placenta and postpubertal testicular Leydig cells, respectively, reflecting distinct functional roles for both hormones within the reproductive tract of Strepsirrhini.
- Mathieu MN et al.
- Synthesis, conformational studies and biological activity of N(alpha)-mono-biotinylated rat relaxin.
- J Pept Res. 2001; 57: 374-82
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Biotin-avidin immobilization can be a useful tool in structure-function studies of hormone receptors. A crucial step is the preparation of a specifically biotinylated hormone that is able to bind to its receptor while leaving the biotin group free for interaction with avidin. The receptor for relaxin, an ovarian peptidic hormone produced during pregnancy, has not yet been isolated. We therefore undertook to prepare a specifically monobiotinylated rat relaxin for use in ligand-searching strategies. Rat relaxin is a convenient analogue because reliable bioassays exist, thus allowing assessment of the effect of N-biotinylation on bioactivity. To help improve the yield of the two-chain, three-disulfide bond rat relaxin, 2-hydroxy-4-methoxybenzyl (Hmb) backbone protection was used during the solid-phase assembly of the B-chain to help prevent any possible chain aggregation. As a final step, while the protected peptide was still on the resin, the biotin label was introduced at the N-terminus of the B-chain using standard coupling protocols. The chain combination with the A-chain was accomplished in reasonable yield. Secondary structural measurements demonstrated that the biotin caused the starting B-chain to adopt a more ordered conformation. The labelled synthetic relaxin exhibited similar circular dichroism spectra to native and synthetic single B-chain peptides. In addition, the biotinylated relaxin showed no significant difference in its chronotropic activity in the rat isolated heart assay compared with the native peptide. Biosensor studies showed that antibody recognition was retained upon attachment of the synthetic relaxin to the streptavidin-derivatized surface.
- Zarreh-Hoshyari-Khah R, Bartsch O, Einspanier A, Pohnke Y, Ivell R
- Bioactivity of recombinant prorelaxin from the marmoset monkey.
- Regul Pept. 2001; 97: 139-46
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The hormone relaxin (RLX) is generally present in the serum of humans and primates as a heterodimer, though some unprocessed prohormone may also be present. In order to test whether this proRLX is biologically relevant for human or primate physiology, recombinant marmoset monkey proRLX was synthesized in a baculovirus-infected cell system and tested in different bioassays. Marmoset proRLX is >70% identical to human H2 proRLX, especially in the so-called receptor-binding region of the B-peptide. The bioassay systems used were (a) cAMP production by human endometrial stromal cells and (b) cAMP production by the human monocyte cell line THP-1. In both bioassay systems recombinant proRLX showed comparable EC(50) values to pure porcine heterodimeric relaxin (porcine relaxin, 1.5-2.0 nM; marmoset prorelaxin 4.0-5.0 nM). Additionally, recombinant marmoset prorelaxin was shown to stimulate steroidogenesis in primary cultures of marmoset ovarian theca cells, though with a lower apparent activity than porcine relaxin. It thus appears that precursor processing of human or primate relaxin is not an essential prerequisite for the acquisition of bioactivity, as it is for the closely related hormone insulin, and that circulating prorelaxin is physiologically relevant.
- Dawson NF et al.
- Solid-phase synthesis of ovine Leydig cell insulin-like peptide--a putative ovine relaxin?
- J Pept Res. 1999; 53: 542-7
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The primary structure of ovine Leydig cell insulin-like peptide (Ley I-L) was recently deduced from the corresponding cDNA sequence. It consists of two peptide chains and three disulphide bonds in an arrangement similar to both relaxin and insulin. As in relaxin B-chain, an Arg-X-X-X-Arg sequence exists within the Ley I-L B-chain although it is located four residues towards the C-terminus from the corresponding position within relaxin. This sequence of amino acids is known to be essential for relaxin biological activity and its presence in Ley I-L suggested that the peptide might possess a relaxin-like function. Ovine Ley I-L was assembled by Fmoc-solid-phase synthesis of the separate chains followed by their combination in solution at high pH. The purity and identity of the chain-combined peptide was confirmed by chemical characterization including mass spectrometry. At physiological concentrations, the peptide was shown not to possess relaxin-like activity in the rat isolated atrial chronotropic and inotropic assay. This strongly suggests that Ley I-L is not a relaxin in the sheep. In order to explore further a possible structural relationship between Ley I-L and relaxin, we prepared a synthetic analogue of ovine Ley I-L containing a single replacement of B-chain residue 12, His, with Arg. This was found to possess significant relaxin-like chronotropic and inotropic activity demonstrating that the tertiary structure of Ley I-L is similar to that of relaxin and highlighting the key requirement for the five-residue sequence, Arg-X-X-X-Arg, to be present in position B12-16 for characteristic relaxin activity.
- Wade JD, Lin F, Talbo G, Otvos L Jr, Tan YY, Tregear GW
- Solid phase synthesis and biological activity of rat relaxin.
- Biomed Pept Proteins Nucleic Acids. 1996; 2: 89-92
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The peptide hormone relaxin was isolated in good yield from the ovaries of the pregnant rodent Rattus rattus using a simplified purification schedule. It was subjected to comprehensive chemical characterization to confirm both its purity and predicted composition. The peptide was also chemically synthesized by the solid phase procedure. The two chains comprising the hormone were each assembled by the Boc-polystyrene method and, following conventional purification, combined in solution to form the single intramolecular and two intermolecular disulfide bonds. Following purification, the synthetic rat relaxin was fully chemically characterized and shown to be indistinguishable from the native peptide including by secondary structure analysis using circular dichroism spectroscopy. The native and synthetic rat relaxins were shown to be equally biologically active in the in vitro rat uterine relaxation assay and had pEC50 values that were comparable to synthetic human H2 relaxin.
- Bode W, Stubbs MT
- Spatial structure of thrombin as a guide to its multiple sites of interaction.
- Semin Thromb Hemost. 1993; 19: 321-33
- Niall HD, Bradshaw RA, Bryant-Greenwood GD
- Relaxin: an insulin-related growth factor.
- Prog Clin Biol Res. 1979; 31: 651-8
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Relaxin is an ovarian peptide that is released just prior to parturition to effect changes in the tissues of the birth canal that aid in the delivery of the fetus. Structural studies established that the two constituent polypeptide chains, composed of 22 and 30 amino acids, are joined by two interchain disulfide bonds with an additional third intrachain bridge. As well as the identical pattern of disulfide bonds, relaxin shows an overall 25% identity with insulin. Furthermore, the sequence of relaxin can be incorporated into the known three-dimensional structure of insulin without significant distortion of the main polypeptide chain backbone. The discovery of the insulin-relatedness of relaxin brings to three the number of growth factors that share a common structural gene precursor with insulin. Nerve growth factor and insulin-like growth factor have already been so identified. Inclusion in this hormone family suggests that the mechanism of action may involve internalization as well as complexation with cell surface receptors of target cells.
- Schwabe C, McDonald JK
- Relaxin: a disulfide homolog of insulin.
- Science. 1977; 197: 914-5
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Relaxin, a peptide hormone responsible for the widening of the birth canal in mammals, has been purified from the ovaries of pregnant hogs. The amino acid sequences of its constituent A and B chains were determined, and the positions of the disulfide cross-links were established. Relaxin was shown to be identical to insulin with respect to its disulfide bond distribution, but significant homology was lacking in other positions. These findings suggest that relaxin and insulin were derived from a common ancestral gene. Since the intrauterine mode of propagation is synonymous with the development of mammals, the genetic distance between insulin and relaxin should therefore permit an estimate of the earliest possible time of commitment of one evolutionary branch to the development of mammals. This event was estimated to have occurred about 5 X 10(8) years ago.
