Insulin capture by an insulin-linked polymorphic region G-quadruplex DNA oligonucleotide.
Insulin capture by a G-quadruplex DNA oligonucleotide containing a two-repeat sequence of the insulin-linked polymorphic region (ILPR) of the human insulin gene promoter region is reported. The immobilized oligonucleotide was demonstrated to capture human insulin from standard solutions and from nuclear extracts of pancreatic cells with high selectivity, using affinity MALDI mass spectrometry and affinity capillary chromatography. Insulin was preferentially captured by the two-repeat ILPR oligonucleotide over another G-quadruplex-forming oligonucleotide, the thrombin-binding aptamer, as well as over a single repeat of the ILPR sequence that is not capable of forming the G-quadruplex architecture. Binding was shown to involve the beta chain of insulin. The discovery raises the possibility that insulin may bind to G-quadruplex DNA formed in the ILPR in vivo and thereby play a role in modulation of insulin gene expression, and it provides a basis for design of insulin analogues to probe this hypothesis. The availability of a DNA ligand to human insulin has analytical importance as well, offering an alternative to antibodies for in vitro or in vivo detection and sensing of insulin as well as its isolation and purification from biological samples.
Connor
Adam C
AC
Department of Chemistry, P. M. Gross Chemical Laboratory, Duke University, Durham, North Carolina 27708, USA.
Frederick
Kimberley A
KA
Morgan
Elizabeth J
EJ
McGown
Linda B
LB
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000737:chemistry; D002846:Chromatography, Affinity; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D054856:G-Quadruplexes; D006801:Humans; D007328:Insulin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010179:Pancreas / Q000737:chemistry; D011401:Promoter Regions, Genetic; D012709:Serum Albumin / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D013917:Thrombin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014020:Tissue Extracts / Q000737:chemistry
D052157:Aptamers, Nucleotide; D007328:Insulin; D009841:Oligonucleotides; D012709:Serum Albumin; D014020:Tissue Extracts; D012822:Silicon Dioxide; D004247:DNA; D013917:Thrombin