Insulin-like Growth Factor 1 Analogs Clicked in the C Domain : Chemical Synthesis and Biological Activities

Human insulin-like growth factor 1 (IGF-1) is a 70 amino acid protein hormone, with key impact on growth, development, and lifespan. The physiological and clinical importance of IGF-1 prompted challenging chemical and biological trials toward the development of its analogs as molecular tools for the IGF-1 receptor (IGF1-R) studies and as new therapeutics. Here, we report a new method for the total chemical synthesis of IGF-1 analogs, which entails the solid-phase synthesis of two IGF-1 precursor chains that is followed by the CuI-catalyzed azide-alkyne cycloaddition ligation and by biomimetic formation of a native pattern of disulfides. The connection of the two IGF-1 precursor chains by the triazole-containing moieties, and variation of its neighboring sequences (Arg36 and Arg37), was tolerated in IGF-1R binding and its activation. These new synthetic IGF-1 analogs are unique examples of disulfide bonds' rich proteins with intra main-chain triazole links. The methodology reported here also presents a convenient synthetic platform for the design and production of new analogs of this important human hormone with non-standard protein modifications

Insulin-Insulin-like Growth Factors Hybrids as Molecular Probes of Hormone : Receptor Binding Specificity

Insulin, insulin-like growth factors 1 and 2 (IGF-1 and -2, respectively), and their receptors (IR and IGF-1R) are the key elements of a complex hormonal system that is essential for the development and functioning of humans. The C and D domains of IGFs (absent in insulin) likely play important roles in the differential binding of IGF-1 and -2 to IGF-1R and to the isoforms of IR (IR-A and IR-B) and specific activation of these receptors. Here, we attempted to probe the impact of IGF-1 and IGF-2 D domains (DI and DII, respectively) and the IGF-2 C domain (CII) on the receptor specificity of these hormones. For this, we made two types of insulin hybrid analogues: (i) with the C-terminus of the insulin A chain extended by the amino acids from the DI and DII domains and (ii) with the C-terminus of the insulin B chain extended by some amino acids derived from the CII domain. The receptor binding affinities of these analogues and their receptor autophosphorylation potentials were characterized. Our results indicate that the DI domain has a more negative impact than the DII domain does on binding to IR, and that the DI domain Pro-Leu-Lys residues are important factors for a different IR-A versus IR-B binding affinity of IGF-1. We also showed that the additions of amino acids that partially "mimic" the CII domain, to the C-terminus of the insulin B chain, change the binding and autophosphorylation specificity of insulin in favor of the "metabolic" IR-B isoform. This opens new venues for rational enhancement of insulin IR-B specificity by modifications beyond the C-terminus of its B chain

Nuclear Charge Radii of Be-7,9,10 and the one-neutron halo nucleus Be-11

Nuclear charge radii of $^{7,9,10,11}$Be have been determined by high-precision laser spectroscopy. On-line measurements were performed with collinear laser spectroscopy in the $2s_{1/2} \to 2p_{1/2}$ transition on a beam of Be$^{+}$ ions. Collinear and anticollinear laser beams were used simultaneously and the absolute frequency determination using a frequency comb yielded an accuracy in the isotope-shift measurements of about 1 MHz. Combination with accurate calculations of the mass-dependent isotope shifts yield nuclear charge radii. The charge radius decreases from $^7$Be to $^{10}$Be and then increases for the halo nucleus $^{11}$Be. When comparing our results with predictions of {\it ab initio} nuclear structure calculations we find good agreement. Additionally, the nuclear magnetic moment of $^7$Be was determined to be $-1.3995(5)\mu_{\rm N}$ and that of $^{11}$Be from a previous $\beta$-NMR measurement was confirmed.Comment: 4 pages, 2 figures calculated mass shift values have been re-evaluated with the latest mass values for the beryllium isotopes and the nuclear polarization contribution for Be-11, published by K. Pachucki et al. ater submission of our manuscript, is also included no

Comparison of Technology and Technical Efficiency in Cereal Production among EU Countries

The paper presents the analysis of cereal production in the EU. The analysis provides the comparison of production technologies and technical efficiency among EU countries using the country specific multiple output distance function models in the first step and metafrontier approach in the second step to determine the level and development of technical efficiency. The results show the high technical efficiency of cereal producers in the analyzed countries. On average, the differences in technical efficiency among the analyzed countries are not pronounced; however, the technologies used as well as the determinants of technical efficiency differ significantly

Identification and Valuation of Public Goods within the Vertical of Cattle Breeding

The paper identifies and discusses the production of public goods in the vertical of cattle breeding. The cattle breeding vertical was divided into four basic levels – producer, processor, retailer, and consumer and main public goods were determined and discussed. Moreover, it provides the methods for the valuation of public goods. The method is applied in the estimation of manure shadow price. Using the fitted multiple output distance function with two market and one non-market output and applying the Lagrange method and the Shephard’s dual lemma the shadow price of manure was calculated. The results show that the shadow prices differ significantly among the groups of farmers. This especially holds for the classification of groups according to the size and technology of production

Toward the insulin-IGF-I intermediate structures: functional and structural properties of the [TyrB25NMePheB26] insulin mutant

The origins of differentiation of insulin from insulin-like growth factor I (IGF-I) are still unknown. To address the problem of a structural and biological switch from the mostly metabolic hormonal activity of insulin to the predominant growth factor activities of IGF-I, an insulin analogue with IGF-I-like structural features has been synthesized. Insulin residues PheB25 and TyrB26 have been swapped with the IGF-I-like Tyr24 and Phe25 sequence with a simultaneous methylation of the peptide nitrogen of residue PheB26. These modifications were expected to introduce a substantial kink in the main chain, as observed at residue Phe25 in the IGF-I crystal structure. These alterations should provide insight into the structural origins of insulin−IGF-I structural and functional divergence. The [TyrB25NMePheB26] mutant has been characterized, and its crystal structure has been determined. Surprisingly, all of these changes are well accommodated within an insulin R6 hexamer. Only one molecule of each dimer in the hexamer responds to the structural alterations, the other remaining very similar to wild-type insulin. All alterations, modest in their scale, cumulate in the C-terminal part of the B-chain (residues B23−B30), which moves toward the core of the insulin molecule and is associated with a significant shift of the A1 helix toward the C-terminus of the B-chain. These changes do not produce the expected bend of the main chain, but the fold of the mutant does reflect some structural characteristics of IGF-1, and in addition establishes the COA19−NHB25 hydrogen bond, which is normally characteristic of T-state insulin