| Chemical synthesis and biological activity of rat INSL3. | |
| | |
MedLine Citation:
|
PMID: 11587188 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The recently identified protein, insulin 3 (INSL3), has structural features that make it a bona fide member of the insulin superfamily. Its predicted amino acid sequence contains the classic two-peptide chain (A- and B-) structure with conserved cysteine residues that results in a disulphide bond disposition identical to that of insulin. Recently, the generation of insl3 knockout mice has demonstrated that testicular descent is blocked due to the failure of a specific ligament, the gubernaculum, to develop. The mechanism by which INSL3 exerts its action on the gubernaculum is currently unknown. The purpose of this study was to, for the first time, synthesize rat INSL3 and test its action on organ cultures of foetal rat gubernaculum. INSL3 also contains a cassette of residues Arg-X-X-X-Arg within the B-chain, a motif that is essential for characteristic activity of another related member of the superfamily, relaxin. Hence, the relaxin activity of rat INSL3 was also tested in two different relaxin bioassays. The primary structure of rat INSL3 was determined by deduction from its cDNA sequence and successfully prepared by solid phase peptide synthesis of the two constituent chains followed by their combination in solution. Following confirmation of its chemical integrity by a variety of analytical techniques, circular dichroism spectroscopy confirmed the presence of high beta-turn and alpha-helical content, with a remarkable spectral similarity to the synthetic ovine INSL3 peptide and to synthetic rat relaxin. The synthetic rat INSL3 bound with very low affinity to rat relaxin receptors and had no activity in a relaxin bioassay. Furthermore, it did not augment or antagonize relaxin activity. The rat INSL3 did however induce growth of foetal rat gubernaculum in whole organ cultures demonstrating that INSL3 has a direct action on this structure. |
| | |
Authors:
|
K J Smith; J D Wade; A A Claasz; L Otvos; C Temelcos; Y Kubota; J M Hutson; G W Tregear; R A Bathgate |
Related Documents
:
|
6461018 - Studies concerning the role of myocardial hypertrophy and of intramural tension as dete... 4722038 - Further evidence for a central hypotensive action of alpha-methyldopa in both the rat a... 14192548 - The rat electroretinogram. ii. bloch's law and the latency mechanism of the b-wave. 13489188 - The effect of diisopropyl fluorophosphonate on the sensitivity of rats to centrally act... 8822138 - Effects of early plasmin digests of fibrinogen on isometric tension development in isol... 3086478 - Thyrotrophin-releasing hormone does not accumulate glycosylated thyrotrophin, but chang... |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
|
Title: Journal of peptide science : an official publication of the European Peptide Society Volume: 7 ISSN: 1075-2617 ISO Abbreviation: J. Pept. Sci. Publication Date: 2001 Sep |
Date Detail:
|
Created Date: 2001-10-05 Completed Date: 2002-03-25 Revised Date: 2006-11-15 |
Medline Journal Info:
|
Nlm Unique ID: 9506309 Medline TA: J Pept Sci Country: England |
Other Details:
|
Languages: eng Pagination: 495-501 Citation Subset: IM |
Affiliation:
|
The Howard Florey Institute, University of Melbourne, Victoria, Australia. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Amino Acid Sequence Animals Biological Assay Circular Dichroism Conserved Sequence Cyclic AMP / metabolism Cysteine / chemistry DNA, Complementary / metabolism Dose-Response Relationship, Drug Insulin Ligands Male Molecular Sequence Data Peptides / chemistry Protein Binding Protein Structure, Tertiary Proteins / chemical synthesis*, metabolism* Rats Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Testis / embryology Time Factors |
| Chemical | |
Reg. No./Substance:
|
0/DNA, Complementary; 0/Leydig insulin-like protein; 0/Ligands; 0/Peptides; 0/Proteins; 11061-68-0/Insulin; 52-90-4/Cysteine; 60-92-4/Cyclic AMP |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Improved preparation of amyloid-beta peptides using DBU as Nalpha-Fmoc deprotection reagent.
Next Document: Synthesis and application of Fmoc-His(3-Bum)-OH.