Abstract
A novel approach to affinity columns is described that is based on the high avidity of biotinylated molecules for avidin attached to solid supports. Biocytin amide [Nepsilon-(+)-biotinyllysine amide] was coupled to the COOH-terminal carboxyl group of corticotropin(1-24) [ACTH(1-24)] to form [biocytin25]ACTH(1-25) amide. The ability of this peptide to stimulate steroidogenesis of bovine adrenocortical cells was within experimental error identical to that of ACTH(1-24). The peptide also binds to avidin and avidin-Sepharose, forming stable complexes. Thus, with biotin as the anchor, the adrenocorticotropically active segment of the ACTH molecule was attached to a solid support in a targeted manner. The general applicability of this principle for the attachment of peptides and proteins to solid supports is discussed.
Full text
PDF![3516](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df53/431147/657b50fa7857/pnas00040-0197.png)
![3517](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df53/431147/8f1c28fabb63/pnas00040-0198.png)
![3518](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df53/431147/f3bbf4a15827/pnas00040-0199.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bodanszky A., Bodanszky M. Sepharose-avidin column for the binding of biotin or biotin-containing peptides. Experientia. 1970 Mar 15;26(3):327–327. doi: 10.1007/BF01900128. [DOI] [PubMed] [Google Scholar]
- Civen M., Selinger R. C. ACTH diazotized to agarose: effects on isolated adrenal cells. Biochem Biophys Res Commun. 1971 May 21;43(4):793–799. doi: 10.1016/0006-291x(71)90686-3. [DOI] [PubMed] [Google Scholar]
- Fauchère J. L., Pelican G. M. Chemisch-spezifische Kopplung von Adrenocorticotropin- und Angiotensin-II-Derivaten an Polymerträger für die Affinitätschromatographie. Helv Chim Acta. 1975 Nov 5;58(7):1984–1994. doi: 10.1002/hlca.19750580713. [DOI] [PubMed] [Google Scholar]
- Finn F. M., Johns P. A., Nishi N., Hofmann K. Differential response to adrenocorticotropic hormone analogs of bovine adrenal plasma membranes and cells. J Biol Chem. 1976 Jun 25;251(12):3576–3585. [PubMed] [Google Scholar]
- Green N. M. Avidin. Adv Protein Chem. 1975;29:85–133. doi: 10.1016/s0065-3233(08)60411-8. [DOI] [PubMed] [Google Scholar]
- Hofmann K., Finn F. M., Limetti M., Montibeller J., Zanetti G. Studies on polypeptides. XXXIV. Enzymic properties of partially synthetic De(16-20)- and De(15-20)-ribonucleases S'1-3. J Am Chem Soc. 1966 Aug 5;88(15):3633–3639. doi: 10.1021/ja00967a030. [DOI] [PubMed] [Google Scholar]
- Hofmann K., Wingender W., Finn F. M. Correlation of adrenocorticotropic activity of ACTH analogs with degree of binding to an adrenal cortical particulate preparation. Proc Natl Acad Sci U S A. 1970 Oct;67(2):829–836. doi: 10.1073/pnas.67.2.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moss J., Lane M. D. The biotin-dependent enzymes. Adv Enzymol Relat Areas Mol Biol. 1971;35:321–442. doi: 10.1002/9780470122808.ch7. [DOI] [PubMed] [Google Scholar]
- Ontjes D. A., Anfinsen C. B. Synthetic studies of structure-function relationships in staphylococcal nuclease. Synthetic analogues of fragment P2. J Biol Chem. 1969 Dec 10;244(23):6316–6322. [PubMed] [Google Scholar]
- Schimmer B. P., Ueda K., Sato G. H. Site of action of adrenocorticotropic hormone (ACTH) in adrenal cell cultures. Biochem Biophys Res Commun. 1968 Sep 6;32(5):806–810. doi: 10.1016/0006-291x(68)90312-4. [DOI] [PubMed] [Google Scholar]
- Seelig S., Sayers G. Isolated adrenal cortex cells: ACTH agonists, partial agonists, antagonists; cyclic AMP and corticosterone production. Arch Biochem Biophys. 1973 Jan;154(1):230–239. doi: 10.1016/0003-9861(73)90053-2. [DOI] [PubMed] [Google Scholar]