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R Gill C Verma B Wallach B Urs? J Pitts A Wollmer P De Meyts S Wood 《Protein engineering》1999,12(4):297-303
Insulin-like growth factor-1 (IGF-1) is a serum protein which unexpectedly folds to yield two stable tertiary structures with different disulphide connectivities; native IGF-1 [18-61,6-48,47-52] and IGF-1 swap [18-61,6-47, 48-52]. Here we demonstrate in detail the biological properties of recombinant human native IGF-1 and IGF-1 swap secreted from Saccharomyces cerevisiae. IGF-1 swap had a approximately 30 fold loss in affinity for the IGF-1 receptor overexpressed on BHK cells compared with native IGF-1.The parallel increase in dose required to induce negative cooperativity together with the parallel loss in mitogenicity in NIH 3T3 cells implies that disruption of the IGF-1 receptor binding interaction rather than restriction of a post-binding conformational change is responsible for the reduction in biological activity of IGF-1 swap. Interestingly, the affinity of IGF-1 swap for the insulin receptor was approximately 200 fold lower than that of native IGF-1 indicating that the binding surface complementary to the insulin receptor (or the ability to attain it) is disturbed to a greater extent than that to the IGF-1 receptor. A 1.0 ns high-temperature molecular dynamics study of the local energy landscape of IGF-1 swap resulted in uncoiling of the first A-region alpha-helix and a rearrangement in the relative orientation of the A- and B-regions. The model of IGF-1 swap is structurally homologous to the NMR structure of insulin swap and CD spectra consistent with the model are presented. However, in the model of IGF-1 swap the C-region has filled the space where the first A-region alpha-helix has uncoiled and this may be hindering interaction of Val44 with the second insulin receptor binding pocket. 相似文献
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From experience in the Northampton/Kettering area 9 to 10% of all patients in a general hospital requiring care in an intensive therapy unit were aged 12 years or under. Fifty-nine children were admitted to the intensive therapy unit of Northampton General Hospital between May 1967 and August 1969. Of these, 22 had been injured in road traffic accidents, five were surgical emergencies, five had meningitis, four status epilepticus, and four respiratory infections.All of the 30 families interviewed were in favour of their child being admitted to the unit, and none considered that the experience had had any lasting adverse psychological effect on the child. It is suggested that certain carefully selected child patients do benefit from the facilities of an intensive therapy unit, and for this reason such units must be designed, equipped, and staffed with this in mind. 相似文献
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P J Davison P Norton S C Wallis L Gill M Cook R Williamson S E Humphries 《Biochemical and biophysical research communications》1986,136(3):876-884
A cDNA probe corresponding to the mRNA sequence for apolipoprotein E (apo E) was used to screen two independently-constructed human genomic libraries. Two recombinants (lambda E-2, and lambda E2-1), isolated using the apo E cDNA probe, also contain part or all of the apo CI gene. Hybridisation studies using both apo E and apo CI cDNA probes show that these two genes are in the same orientation and separated by 4 kb. 相似文献
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