The Experts below are selected from a list of 2358 Experts worldwide ranked by ideXlab platform
Christopher R. Parish - One of the best experts on this subject based on the ideXlab platform.
-
use of sulfated linked Cyclitols as heparan sulfate mimetics to probe the heparin heparan sulfate binding specificity of proteins
Journal of Biological Chemistry, 2005Co-Authors: Craig Freeman, Vito Ferro, Martin G. Banwell, Ligong Liu, Kathryn J. Brown, Anna Bezos, Christopher R. ParishAbstract:Heparin and heparan sulfate (HS) are structurally diverse glycosaminoglycans (GAG) that are known to interact, via unique structural motifs, with a wide range of functionally distinct proteins and modulate their biological activity. To define the GAG motifs that interact with proteins, we assessed the ability of 15 totally synthetic HS mimetics to interact with 10 functionally diverse proteins that bind heparin/HS. The HS mimetics consisted of cyclitol-based pseudo-sugars coupled by linkers of variable chain length, flexibility, orientation, and hydrophobicity, with variations in sulfation also being introduced into some molecules. Three of the proteins tested, namely hepatocyte growth factor, eotaxin, and elastase, failed to interact with any of the sulfated linked Cyclitols. In contrast, each of the remaining seven proteins tested exhibited a unique reactivity pattern with the panel of HS mimetics, with tetrameric Cyclitols linked by different length alkyl chains being particularly informative. Thus, compounds with short alkyl spacers (2–3 carbon atoms) effectively blocked the interaction of fibroblast growth factor-1 (FGF-1) and lipoprotein lipase with heparin/HS, whereas longer chain spacers (7–10 carbon atoms) were required for optimal inhibition of FGF-2 and vascular endothelial growth factor binding. This effect was most pronounced with the chemokine, interleukin-8, where alkyl-linked tetrameric Cyclitols were essentially inactive unless a spacer of >7 carbon atoms was used. The heparin-inhibitable enzymes heparanase and cathepsin G also displayed characteristic inhibition patterns, cathepsin G interacting promiscuously with most of the sulfated Cyclitols but heparanase activity being inhibited most effectively by HS mimetics that structurally resemble a sulfated pentasaccharide. These data indicate that a simple panel of HS mimetics can be used to probe the HS binding specificity of proteins, with the position of anionic groups in the HS mimetics being critical.
-
Use of Sulfated Linked Cyclitols as Heparan Sulfate Mimetics to Probe the Heparin/Heparan Sulfate Binding Specificity of Proteins
The Journal of biological chemistry, 2005Co-Authors: Craig Freeman, Vito Ferro, Martin G. Banwell, Ligong Liu, Kathryn J. Brown, Anna Bezos, Christopher R. ParishAbstract:Heparin and heparan sulfate (HS) are structurally diverse glycosaminoglycans (GAG) that are known to interact, via unique structural motifs, with a wide range of functionally distinct proteins and modulate their biological activity. To define the GAG motifs that interact with proteins, we assessed the ability of 15 totally synthetic HS mimetics to interact with 10 functionally diverse proteins that bind heparin/HS. The HS mimetics consisted of cyclitol-based pseudo-sugars coupled by linkers of variable chain length, flexibility, orientation, and hydrophobicity, with variations in sulfation also being introduced into some molecules. Three of the proteins tested, namely hepatocyte growth factor, eotaxin, and elastase, failed to interact with any of the sulfated linked Cyclitols. In contrast, each of the remaining seven proteins tested exhibited a unique reactivity pattern with the panel of HS mimetics, with tetrameric Cyclitols linked by different length alkyl chains being particularly informative. Thus, compounds with short alkyl spacers (2–3 carbon atoms) effectively blocked the interaction of fibroblast growth factor-1 (FGF-1) and lipoprotein lipase with heparin/HS, whereas longer chain spacers (7–10 carbon atoms) were required for optimal inhibition of FGF-2 and vascular endothelial growth factor binding. This effect was most pronounced with the chemokine, interleukin-8, where alkyl-linked tetrameric Cyclitols were essentially inactive unless a spacer of >7 carbon atoms was used. The heparin-inhibitable enzymes heparanase and cathepsin G also displayed characteristic inhibition patterns, cathepsin G interacting promiscuously with most of the sulfated Cyclitols but heparanase activity being inhibited most effectively by HS mimetics that structurally resemble a sulfated pentasaccharide. These data indicate that a simple panel of HS mimetics can be used to probe the HS binding specificity of proteins, with the position of anionic groups in the HS mimetics being critical.
Marianne Popp - One of the best experts on this subject based on the ideXlab platform.
-
Cyclitols protect glutamine synthetase and malate dehydrogenase against heat induced deactivation and thermal denaturation
Biochemical and Biophysical Research Communications, 2006Co-Authors: Martina Jaindl, Marianne PoppAbstract:The accumulation of Cyclitols in plants is a widespread response that provides protection against various environmental stresses. The capacity of myo-Inositol, pinitol, quercitol, and other compatible solutes (i.e., sorbitol, proline, and glycinebetaine) to protect proteins against thermally induced denaturation and deactivation was examined. Enzymatic activity measurements of L-glutamine synthetase from Escherichia coli and Hordeum vulgare showed that the presence of Cyclitols during heat treatment resulted in a significantly higher percentage of residual activity. CD spectroscopy experiments were used to study thermal stabilities of protein secondary structures upon the addition of myo-Inositol, pinitol, and glucose. 0.4 M myo-Inositol was observed to raise the melting temperature (Tm) of GS from E. coli by 3.9 degrees C and MDH from pig heart by 3.4 degrees C, respectively. Pinitol showed an increase in Tm of MDH by 3.8 degrees C, whereas glucose was not effective. Our results show a great potential of stabilizing proteins by the addition of Cyclitols.
-
Cyclitols protect glutamine synthetase and malate dehydrogenase against heat induced deactivation and thermal denaturation.
Biochemical and biophysical research communications, 2006Co-Authors: Martina Jaindl, Marianne PoppAbstract:Abstract The accumulation of Cyclitols in plants is a widespread response that provides protection against various environmental stresses. The capacity of myo -Inositol, pinitol, quercitol, and other compatible solutes (i.e., sorbitol, proline, and glycinebetaine) to protect proteins against thermally induced denaturation and deactivation was examined. Enzymatic activity measurements of l -glutamine synthetase from Escherichia coli and Hordeum vulgare showed that the presence of Cyclitols during heat treatment resulted in a significantly higher percentage of residual activity. CD spectroscopy experiments were used to study thermal stabilities of protein secondary structures upon the addition of myo -Inositol, pinitol, and glucose. 0.4 M myo -Inositol was observed to raise the melting temperature ( T m ) of GS from E. coli by 3.9 °C and MDH from pig heart by 3.4 °C, respectively. Pinitol showed an increase in T m of MDH by 3.8 °C, whereas glucose was not effective. Our results show a great potential of stabilizing proteins by the addition of Cyclitols.
-
Cyclitols as cryoprotectants for spinach and chickpea thylakoids
Environmental and experimental botany, 2000Co-Authors: Birgit Orthen, Marianne PoppAbstract:Abstract Thylakoid membranes isolated from either spinach or chickpea leaves were used as a model system for evaluating the capacity of Cyclitols to act as cryoprotectants. The effect of freezing for 3 h at −18°C on cyclic photophosphorylation and electron transport was measured. The Cyclitols, ononitol, O -methyl- muco -inositol, pinitol, quebrachitol and quercitol at 50–150 mol m −3 decreased membrane damage by freezing and thawing to a similar degree as the well known cryoprotectants sucrose and trehalose. On addition of the cryotoxic solute NaCl (100 mol m −3 ) to the test system these methylated cyclohexanhexols again provided a protection comparable to that of the two disaccharides. Quercitol (cyclohexanpentol) was not effective when added in lower concentrations (50–100 mol m −3 ) and in case of this cyclitol a ratio of membrane toxic to membrane compatible solute of 0.66 was apparently needed to prevent a loss of cyclic photophosphorylation. Little difference was observed in the results from spinach or chickpea thylakoids although these plants naturally accumulate different cyto-solutes (spinach: glycinebetaine; chickpea: pinitol).
-
Hydroxyl radical scavenging properties of Cyclitols
Proceedings of the Royal Society of Edinburgh. Section B. Biological Sciences, 1994Co-Authors: Birgit Orthen, Marianne Popp, Nicholas SmirnoffAbstract:Cyclitols are low molecular weight substances which accumulate in plant cells in response to various environmental stress situations, for example drought (Ford 1984), salinity (Gorham et al . 1984), low temperature (Richter et al . 1990). Apart from their more general role in osmotic adjustment, only in the case of salt stress is their mode of function well understood. Cyclitols (e.g. pinitol) accumulate when plants are exposed to increasing salt concentration (Paul & Cockburn 1989) and act as compatible solutes (Sommer et al . 1990) as defined by Brown & Simpson (1972). The significance of cyclitol accumulation in stress adaptation of plants to drought and cold still remains uncertain. However, it is generally accepted that drought and cold as well as several other stress situations lead to an enhanced generation of oxygen free radicals (Elstner 1990; Smirnoff & Colombe 1988), including the hydroxyl radical as the most harmful one. The report by Smirnoff & Cumbes (1989) that myo-inositol is an effective hydroxyl radical scavenger prompted us to test other naturally-occuring Cyclitols like pinitol, quebrachitol, 1-D-1-O-methyl-muco-inositol, ononitol and quercitol for their ability to scavenge hydroxyl radicals.
-
The physiological importance of accumulation of Cyclitols in Viscum album L.
New Phytologist, 1992Co-Authors: Andreas Richter, Marianne PoppAbstract:SUMMARY Investigations of Viscum album on 16 different host species revealed an endogenous cyclitol pattern with pinitol, 1-O-methyl-muco-inositol, quebrachitol, chiro-inositol, an unidentified O-methyl-inositol and traces of ononitol. Host-specific Cyclitols including bornesitol, quercitol, viburnitol, scyllo-inositol and the hexitol, sorbitol, were also stored in the mistletoe. The endogenous Cyclitols were accumulated to such high concentrations that they made a large contribution to the osmotic potential. It was estimated that 22.6–43 % of mistletoe carbon is derived from the host.
Craig Freeman - One of the best experts on this subject based on the ideXlab platform.
-
use of sulfated linked Cyclitols as heparan sulfate mimetics to probe the heparin heparan sulfate binding specificity of proteins
Journal of Biological Chemistry, 2005Co-Authors: Craig Freeman, Vito Ferro, Martin G. Banwell, Ligong Liu, Kathryn J. Brown, Anna Bezos, Christopher R. ParishAbstract:Heparin and heparan sulfate (HS) are structurally diverse glycosaminoglycans (GAG) that are known to interact, via unique structural motifs, with a wide range of functionally distinct proteins and modulate their biological activity. To define the GAG motifs that interact with proteins, we assessed the ability of 15 totally synthetic HS mimetics to interact with 10 functionally diverse proteins that bind heparin/HS. The HS mimetics consisted of cyclitol-based pseudo-sugars coupled by linkers of variable chain length, flexibility, orientation, and hydrophobicity, with variations in sulfation also being introduced into some molecules. Three of the proteins tested, namely hepatocyte growth factor, eotaxin, and elastase, failed to interact with any of the sulfated linked Cyclitols. In contrast, each of the remaining seven proteins tested exhibited a unique reactivity pattern with the panel of HS mimetics, with tetrameric Cyclitols linked by different length alkyl chains being particularly informative. Thus, compounds with short alkyl spacers (2–3 carbon atoms) effectively blocked the interaction of fibroblast growth factor-1 (FGF-1) and lipoprotein lipase with heparin/HS, whereas longer chain spacers (7–10 carbon atoms) were required for optimal inhibition of FGF-2 and vascular endothelial growth factor binding. This effect was most pronounced with the chemokine, interleukin-8, where alkyl-linked tetrameric Cyclitols were essentially inactive unless a spacer of >7 carbon atoms was used. The heparin-inhibitable enzymes heparanase and cathepsin G also displayed characteristic inhibition patterns, cathepsin G interacting promiscuously with most of the sulfated Cyclitols but heparanase activity being inhibited most effectively by HS mimetics that structurally resemble a sulfated pentasaccharide. These data indicate that a simple panel of HS mimetics can be used to probe the HS binding specificity of proteins, with the position of anionic groups in the HS mimetics being critical.
-
Use of Sulfated Linked Cyclitols as Heparan Sulfate Mimetics to Probe the Heparin/Heparan Sulfate Binding Specificity of Proteins
The Journal of biological chemistry, 2005Co-Authors: Craig Freeman, Vito Ferro, Martin G. Banwell, Ligong Liu, Kathryn J. Brown, Anna Bezos, Christopher R. ParishAbstract:Heparin and heparan sulfate (HS) are structurally diverse glycosaminoglycans (GAG) that are known to interact, via unique structural motifs, with a wide range of functionally distinct proteins and modulate their biological activity. To define the GAG motifs that interact with proteins, we assessed the ability of 15 totally synthetic HS mimetics to interact with 10 functionally diverse proteins that bind heparin/HS. The HS mimetics consisted of cyclitol-based pseudo-sugars coupled by linkers of variable chain length, flexibility, orientation, and hydrophobicity, with variations in sulfation also being introduced into some molecules. Three of the proteins tested, namely hepatocyte growth factor, eotaxin, and elastase, failed to interact with any of the sulfated linked Cyclitols. In contrast, each of the remaining seven proteins tested exhibited a unique reactivity pattern with the panel of HS mimetics, with tetrameric Cyclitols linked by different length alkyl chains being particularly informative. Thus, compounds with short alkyl spacers (2–3 carbon atoms) effectively blocked the interaction of fibroblast growth factor-1 (FGF-1) and lipoprotein lipase with heparin/HS, whereas longer chain spacers (7–10 carbon atoms) were required for optimal inhibition of FGF-2 and vascular endothelial growth factor binding. This effect was most pronounced with the chemokine, interleukin-8, where alkyl-linked tetrameric Cyclitols were essentially inactive unless a spacer of >7 carbon atoms was used. The heparin-inhibitable enzymes heparanase and cathepsin G also displayed characteristic inhibition patterns, cathepsin G interacting promiscuously with most of the sulfated Cyclitols but heparanase activity being inhibited most effectively by HS mimetics that structurally resemble a sulfated pentasaccharide. These data indicate that a simple panel of HS mimetics can be used to probe the HS binding specificity of proteins, with the position of anionic groups in the HS mimetics being critical.
Bogusław Buszewski - One of the best experts on this subject based on the ideXlab platform.
-
New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS
International journal of molecular sciences, 2020Co-Authors: Gulyaim N. Sagandykova, Paweł Pomastowski, Justyna Walczak-skierska, Fernanda Monedeiro, Bogusław BuszewskiAbstract:A combination of electrochemistry (EC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (off-line EC-MALDI-TOF-MS) was applied for determination of the studied biologically active compounds (D-glucose, D-fructose, D-galactose, D-pinitol, L-chiro-inositol, and myo-inositol) and their possible electrochemical metabolites. In this work, boron-doped diamond electrode (BDD) was used as a working electrode. MALDI-TOF-MS experiments were carried out (both in positive and negative ion modes and using two matrices) to identify the structures of electrochemical products. This was one of the first applications of the EC system for the generation of electrochemical products produced from saccharides and Cyclitols. Moreover, exploratory data analysis approaches (correlation networks, hierarchical cluster analysis, weighted plots) were used in order to present differences/similarities between the obtained spectra, regarding the class of analyzed compounds, ionization modes, and used matrices. This work presents the investigation and comparison of fragmentation patterns of sugars, Cyclitols, and their respective products generated through the electrochemistry (EC) process.
-
Correlation Study of Honey Regarding their Physicochemical Properties and Sugars and Cyclitols Content.
Molecules (Basel Switzerland), 2019Co-Authors: Ileana-andreea Rațiu, Hossam Al-suod, Magdalena Ligor, Malgorzata Bukowska, Bogusław BuszewskiAbstract:Honey is a natural sweetener, with an osmotic effect on microorganisms due to the increased sugar content and low amount of water. Cyclitols are minor constituents of honey. They play a defensive role in plants against unfavorable environmental conditions. Honey's physicochemical properties can vary, resulting in a wide range of colors, flavors, scents, antioxidant activity, dissimilar values of pH, acidity, electrical conductivity, etc. Some literature regarding correlation between honey types is already available, but a comprehensive study displaying an ample evaluation of multifarious aspects is still needed. This study focuses on the correlation between 18 honey types, originating from 10 countries, collected during four years, summarizing a total of 38 samples. A total of 6 physicochemical properties and 18 target components (sugars and Cyclitols) were considered as variables. A correlation analysis is presented between the investigated parameters and between honey types, together with the statistical analysis which allowed for observation of the clusters' distribution according with the investigated variables.
-
Supercritical fluid extraction in isolation of Cyclitols and sugars from chamomile flowers.
Journal of separation science, 2019Co-Authors: Hossam Al-suod, Lesław B. Lahuta, Ryszard J. Górecki, Ileana-andreea Rațiu, Aneta Krakowska‐sieprawska, Bogusław BuszewskiAbstract:The aim of the present study was to develop an optimization procedure for supercritical fluid extraction parameters, in order to obtain the highest possible yield of sugars and Cyclitols from plant material. Response surface methodology based on Box-Behnken design was applied to evaluate the effect of: temperature (40, 60, 80°C), pressure (100, 200, 300 bar), and co-solvent (methanol) percentage (20, 25, 30%). As a result of the optimization process, we found that the highest amount of sugars (15.02 mg/gof dried material) and Cyclitols (0.86 mg/g of dried material) was obtained when the following parameters were applied: 80°C, 228 bar, and 30% of methanol. Moreover, co-solvent concentration and temperature had a higher influence onto the obtained amounts compared with the pressure.
-
Simultaneous Determination of Cyclitols and Sugars Following a Comprehensive Investigation of 40 Plants
Food Analytical Methods, 2019Co-Authors: Ileana-andreea Rațiu, Hossam Al-suod, Magdalena Ligor, Tomasz Ligor, Aneta Krakowska, Ryszard Górecki, Bogusław BuszewskiAbstract:Due to the important features of widely unexplored Cyclitols, a comprehensive qualitative and quantitative study is needed. Moreover, measuring the possible available amounts of identified components in plant material represents a stringent need, due to their importance in phytomedicine and their use in food. The purpose of this study was to realize an extended investigation mainly of Cyclitols, but of sugars and sugar alcohols as well, from natural sources. Thus, 17 target compounds (7 Cyclitols and 2 sugar alcohols and 8 sugars) extracted from medicinal and edible plants are reported. All detected components were simultaneous separated in just one chromatographic run, using a single GC column. A number of 52 sources coming from 40 species were studied. Thus, we report 37 new sources of Cyclitols. Moreover, almost for all Cyclitols, the richest source was not investigated previously. Therefore, the obtained results can represent a valuable material for food, pharmaceutical, medical, or cosmetic industry interested in the use of Cyclitols.
-
Pressurized liquid extraction of Cyclitols and sugars: optimization of extraction parameters and selective separation.
Journal of separation science, 2019Co-Authors: Hossam Al-suod, Ryszard J. Górecki, Ileana-andreea Rațiu, Bogusław BuszewskiAbstract:Cyclitols and sugars were obtained as a mixture from Medicago sativa L., in a comparative study by using maceration, and pressurized liquid extraction, as a modern and green extraction techniques. The influence of extraction parameters including: extraction temperature, time and number of cycles on the content of sugars and Cyclitols was investigated based on response surface methodology. The highest total amount of sugars and Cyclitols (62.27 ± 2.30 and 50.35 ± 0.77 mg/g of dry material, respectively) was obtained when extraction was performed at 88°C, for 22 min, in two cycles. The methodology used involved extraction, purification, selective separation (using yeast and anion exchange resin) and derivatization, followed by gas chromatography -mass spectrometry analysis. The use of yeast treatment realized an effective fractionation of Cyclitols and sugars, which allowed the removal of most sugars. The involvement of anion exchange resin after yeast allowed the removal of sugar alcohols and lactose, together with other sugar traces remained and to obtain a solution containing six Cyclitols. The recrystallization of dry residue after solvent evaporation, from ethanol, allowed us to obtain 14.65 mg of white pure crystals identified with NMR spectroscopy, liquid chromatography with mass spectrometry, gas chromatography with mass spectrometry, optical rotation and melting point as analysis D-pinitol.
Vito Ferro - One of the best experts on this subject based on the ideXlab platform.
-
Anti-herpes simplex virus activities of two novel disulphated Cyclitols.
Antiviral Chemistry and Chemotherapy, 2006Co-Authors: Maria Ekblad, Tomas Bergström, Vito Ferro, Martin G. Banwell, Muriel Bonnet, Jens Renner, Edward TrybalaAbstract:By screening a library of sulphated compounds of low molecular weight, we have found that several cyclitol derivatives, each modified with two sulphate groups in addition to pyrrole and various aromatic moieties, inhibited infectivity of herpes simplex virus (HSV) at concentrations approximately 100 times lower than those toxic for cultured cells. These disulphated Cyclitols interfered with HSV-1 attachment to cells, and efficiently reduced the cell-to-cell spread of the virus. This effect is most likely due to their low molecular weight and associated with the compounds' capability to access the narrow intercellular spaces. Furthermore, these disulphated Cyclitols also inactivated infectivity of HSV. However, the virus-inactivating activities of these compounds were to some extent diminished in the presence of human cervical secretions or other protein-rich solutions suggesting that disulphated Cyclitols may have some features of surfactant-type virucides. In conclusion, this new class of anti-HSV compounds offers potential for further development.
-
use of sulfated linked Cyclitols as heparan sulfate mimetics to probe the heparin heparan sulfate binding specificity of proteins
Journal of Biological Chemistry, 2005Co-Authors: Craig Freeman, Vito Ferro, Martin G. Banwell, Ligong Liu, Kathryn J. Brown, Anna Bezos, Christopher R. ParishAbstract:Heparin and heparan sulfate (HS) are structurally diverse glycosaminoglycans (GAG) that are known to interact, via unique structural motifs, with a wide range of functionally distinct proteins and modulate their biological activity. To define the GAG motifs that interact with proteins, we assessed the ability of 15 totally synthetic HS mimetics to interact with 10 functionally diverse proteins that bind heparin/HS. The HS mimetics consisted of cyclitol-based pseudo-sugars coupled by linkers of variable chain length, flexibility, orientation, and hydrophobicity, with variations in sulfation also being introduced into some molecules. Three of the proteins tested, namely hepatocyte growth factor, eotaxin, and elastase, failed to interact with any of the sulfated linked Cyclitols. In contrast, each of the remaining seven proteins tested exhibited a unique reactivity pattern with the panel of HS mimetics, with tetrameric Cyclitols linked by different length alkyl chains being particularly informative. Thus, compounds with short alkyl spacers (2–3 carbon atoms) effectively blocked the interaction of fibroblast growth factor-1 (FGF-1) and lipoprotein lipase with heparin/HS, whereas longer chain spacers (7–10 carbon atoms) were required for optimal inhibition of FGF-2 and vascular endothelial growth factor binding. This effect was most pronounced with the chemokine, interleukin-8, where alkyl-linked tetrameric Cyclitols were essentially inactive unless a spacer of >7 carbon atoms was used. The heparin-inhibitable enzymes heparanase and cathepsin G also displayed characteristic inhibition patterns, cathepsin G interacting promiscuously with most of the sulfated Cyclitols but heparanase activity being inhibited most effectively by HS mimetics that structurally resemble a sulfated pentasaccharide. These data indicate that a simple panel of HS mimetics can be used to probe the HS binding specificity of proteins, with the position of anionic groups in the HS mimetics being critical.
-
Use of Sulfated Linked Cyclitols as Heparan Sulfate Mimetics to Probe the Heparin/Heparan Sulfate Binding Specificity of Proteins
The Journal of biological chemistry, 2005Co-Authors: Craig Freeman, Vito Ferro, Martin G. Banwell, Ligong Liu, Kathryn J. Brown, Anna Bezos, Christopher R. ParishAbstract:Heparin and heparan sulfate (HS) are structurally diverse glycosaminoglycans (GAG) that are known to interact, via unique structural motifs, with a wide range of functionally distinct proteins and modulate their biological activity. To define the GAG motifs that interact with proteins, we assessed the ability of 15 totally synthetic HS mimetics to interact with 10 functionally diverse proteins that bind heparin/HS. The HS mimetics consisted of cyclitol-based pseudo-sugars coupled by linkers of variable chain length, flexibility, orientation, and hydrophobicity, with variations in sulfation also being introduced into some molecules. Three of the proteins tested, namely hepatocyte growth factor, eotaxin, and elastase, failed to interact with any of the sulfated linked Cyclitols. In contrast, each of the remaining seven proteins tested exhibited a unique reactivity pattern with the panel of HS mimetics, with tetrameric Cyclitols linked by different length alkyl chains being particularly informative. Thus, compounds with short alkyl spacers (2–3 carbon atoms) effectively blocked the interaction of fibroblast growth factor-1 (FGF-1) and lipoprotein lipase with heparin/HS, whereas longer chain spacers (7–10 carbon atoms) were required for optimal inhibition of FGF-2 and vascular endothelial growth factor binding. This effect was most pronounced with the chemokine, interleukin-8, where alkyl-linked tetrameric Cyclitols were essentially inactive unless a spacer of >7 carbon atoms was used. The heparin-inhibitable enzymes heparanase and cathepsin G also displayed characteristic inhibition patterns, cathepsin G interacting promiscuously with most of the sulfated Cyclitols but heparanase activity being inhibited most effectively by HS mimetics that structurally resemble a sulfated pentasaccharide. These data indicate that a simple panel of HS mimetics can be used to probe the HS binding specificity of proteins, with the position of anionic groups in the HS mimetics being critical.
