Chelating peptide-immobilized metal ion affinity chromatography. A new concept in affinity chromatography for recombinant proteins - PubMed (original) (raw)
. 1988 May 25;263(15):7211-5.
Affiliations
- PMID: 3284883
Free article
Chelating peptide-immobilized metal ion affinity chromatography. A new concept in affinity chromatography for recombinant proteins
M C Smith et al. J Biol Chem. 1988.
Free article
Abstract
We report our experimental results supporting the hypothesis that a specific metal-chelating peptide (CP) on the NH2 terminus of a protein can be used to purify that protein using immobilized metal ion affinity chromatography (IMAC). The potential utility of this approach resides with recombinant proteins since the nucleotide sequence that codes for the protein can be extended to include codons for the chelating peptide and thereby generate the gene for a chimeric CP-protein that can be cloned, expressed, and affinity-purified with immobilized metal ions. The chelating peptide purification handle could then be removed chemically or enzymatically after purification has been achieved to generate a protein with the natural amino acid sequence. The feasibility of using a chelating peptide as a purification handle has been demonstrated using a leuteinizing hormone-releasing hormone (LHRH) analog, 2-10 LHRH, which contains the previously identified chelating peptide, His-Trp, on the NH2 terminus. 2-10 LHRH had a high affinity for a Ni(II) IMAC column due to the NH2-terminal dipeptide sequence His-Trp, forming a coordination complex with Ni(II), whereas the controls, 3-10 LHRH and 4-10 LHRH, lacking the CP sequence, did not bind. Furthermore, 2-10 LHRH could be purified from a mixture of histidine-containing peptides on a Ni(II) IMAC column in one step. His-Trp proinsulin was used as a model of a recombinant CP-protein. The S-sulfonates of His-Trp-proinsulin and proinsulin were isolated from Escherichia coli engineered to overproduce these proteins as trpLE' fusion proteins. His-Trp-proinsulin(SSO3-)6 had a higher affinity for immobilized Ni(II) than proinsulin (SSO3-)6. Both proteins were eluted by decreasing the pH or by introducing a displacing ligand into the buffer. Ni(II) eluted from the column with much higher concentrations of displacing ligand than the proteins.
Similar articles
- Metal affinity engineering of proinsulin carrying genetically attached (His)10-X-Met affinity tail and removal of the tag by cyanogen bromide.
Ko JH, Chung WJ, Koh S, Park BC, Kwon ST, Kim CH, Lee DS. Ko JH, et al. Biosci Biotechnol Biochem. 1994 Sep;58(9):1694-9. doi: 10.1271/bbb.58.1694. Biosci Biotechnol Biochem. 1994. PMID: 7765485 - Immobilized palladium(II) ion affinity chromatography for recovery of recombinant proteins with peptide tags containing histidine and cysteine.
Kikot P, Polat A, Achilli E, Fernandez Lahore M, Grasselli M. Kikot P, et al. J Mol Recognit. 2014 Nov;27(11):659-68. doi: 10.1002/jmr.2389. J Mol Recognit. 2014. PMID: 25277090 - Perspectives of immobilized-metal affinity chromatography.
Gaberc-Porekar V, Menart V. Gaberc-Porekar V, et al. J Biochem Biophys Methods. 2001 Oct 30;49(1-3):335-60. doi: 10.1016/s0165-022x(01)00207-x. J Biochem Biophys Methods. 2001. PMID: 11694288 Review. - [A new trend toward using metal chelates in affinity chromatography of proteins (review)].
Lopatin SA, Varlamov VP. Lopatin SA, et al. Prikl Biokhim Mikrobiol. 1995 May-Jun;31(3):259-66. Prikl Biokhim Mikrobiol. 1995. PMID: 7638155 Review. Russian.
Cited by
- A Histidine-Rich Extensin from Zea mays Is an Arabinogalactan Protein.
Kieliszewski MJ, Kamyab A, Leykam JF, Lamport DT. Kieliszewski MJ, et al. Plant Physiol. 1992 Jun;99(2):538-47. doi: 10.1104/pp.99.2.538. Plant Physiol. 1992. PMID: 16668920 Free PMC article. - Cold shock induction of recombinant Arctic environmental genes.
Bjerga GE, Williamson AK. Bjerga GE, et al. BMC Biotechnol. 2015 Aug 19;15:78. doi: 10.1186/s12896-015-0185-1. BMC Biotechnol. 2015. PMID: 26286037 Free PMC article. - A method for prediction of the locations of linker regions within large multifunctional proteins, and application to a type I polyketide synthase.
Udwary DW, Merski M, Townsend CA. Udwary DW, et al. J Mol Biol. 2002 Oct 25;323(3):585-98. doi: 10.1016/s0022-2836(02)00972-5. J Mol Biol. 2002. PMID: 12381311 Free PMC article. - Purification of BmR1 recombinant protein.
Arifin N, Basuni M, Lan CA, Yahya AR, Noordin R. Arifin N, et al. Protein J. 2010 Oct;29(7):509-15. doi: 10.1007/s10930-010-9281-1. Protein J. 2010. PMID: 20845068 - Improvement of the crystallizability and expression of an RNA crystallization chaperone.
Ravindran PP, Héroux A, Ye JD. Ravindran PP, et al. J Biochem. 2011 Nov;150(5):535-43. doi: 10.1093/jb/mvr093. Epub 2011 Jul 23. J Biochem. 2011. PMID: 21785128 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous