Linked Life Data

2176911

Source:http://linkedlifedata.com/resource/pubmed/id/2176911

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1991-2-28
pubmed:abstractText
A sensitive binding assay was developed to determine binding characteristics of commercially available [125I-Tyr10]human growth hormone-releasing factor (hGRF) (1-44)NH2 in rat pituitary using 0.1 gland homogenate (70-75 micrograms protein) per incubation tube. Under standard assay conditions, addition of 5 mM EDTA efficiently prevented the degradation of both human and rat GRF for at least 3 h. Association of the ligand was time-dependent: equilibrium was reached within 30 min of incubation at 23 degrees C and remained stable for an additional 150 min (K1 = 5.01 +/- 0.86 nM-1.min-1). Specific binding increased linearly with the amount of protein present in the assay, from 15 to 170 micrograms per incubation tube. This binding was reversible, dissociation occurring almost completely after a 120-min period (K-1 = 8.13 +/- 0.29 x 10(-3) min-1). A concentration of 5-10 mM Mg2+ was required for optimal specific binding whereas 50 mM Mg2+ or monovalent cations such as Na+, K+, Li+ decreased it. Scatchard analysis of cold saturation studies by the Ligand program statistically revealed the presence of two distinct classes of binding sites; the first was of high affinity (0.68 +/- 0.11 nM) and low capacity (140 +/- 22 fmol/pituitary), the second was of lower affinity (590 +/- 347 nM) and higher capacity (38.7 +/- 18.7 pmol/pituitary). Similar values were obtained with various bovine serum albumin (BSA) concentrations and when using crude or washed pituitary homogenates, suggesting that the second low affinity site was not BSA or a soluble protein from the homogenate.(ABSTRACT TRUNCATED AT 250 WORDS)
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
http://linkedlifedata.com/resource/pubmed/chemical/Cations, http://linkedlifedata.com/resource/pubmed/chemical/Edetic Acid, http://linkedlifedata.com/resource/pubmed/chemical/Growth Hormone-Releasing Hormone, http://linkedlifedata.com/resource/pubmed/chemical/Guanine Nucleotides, http://linkedlifedata.com/resource/pubmed/chemical/Iodine Radioisotopes, http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Neuropeptide, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Neurotransmitter, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Pituitary..., http://linkedlifedata.com/resource/pubmed/chemical/Sermorelin, http://linkedlifedata.com/resource/pubmed/chemical/Serum Albumin, Bovine, http://linkedlifedata.com/resource/pubmed/chemical/somatotropin releasing hormone..., http://linkedlifedata.com/resource/pubmed/chemical/somatotropin releasing hormone...
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-8993
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
528
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
291-9
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1990
pubmed:articleTitle
Characterization of [125I-Tyr10]human growth hormone-releasing factor (1-44) amide binding to rat pituitary: evidence for high and low affinity classes of sites.
pubmed:affiliation
Neuroendocrinology Laboratory, Notre-Dame Hospital Research Center, University of Montreal, Que., Canada.
pubmed:publicationType
Journal Article, In Vitro, Research Support, Non-U.S. Gov't