Biomedical subjects

Andrzej J Kulik

Publications and source records attributed to Andrzej J Kulik.

6 recordsLinked to original sources

PubMed2006

Lectin-carbohydrate affinity measured using a quartz crystal microbalance.

The association of two molecules is described by two parameters, association equilibrium and association rate constants, which are characteristic for a given type of interaction. Usually, they are determined for interacting molecules dissolved in solution. However, for many applications one type of molecules is immobilized on a substrate, which may influence the binding kinetics. The studied complex of concanavalin A and carboxypeptidase Y belongs to the lectin-carbohydrate type of interaction involving the recognition of oligosaccharide moieties. The concanavalin A was immobilized on a gold electrode of quartz crystal, while carboxypeptidase Y was added to a buffer (Tris-buffered saline). The constants describing the association of the investigated molecules were determined on the basis of measurements performed using a quartz crystal microbalance in liquid. The obtained values were (0.59+/-0.01)x10(6) M(-1) for the association equilibrium constant and (5.6+/-0.1)x10(4) M(-1)s(-1) for the association rate constant. The saturation binding experiment gave another value of the association constant, (2.7+/-0.02)x10(6) M(-1). The comparison of obtained values with previously published ones verifies that the molecule orientation and binding site accessibility for specific ligands could influence the association equilibrium constant value. The presented measurements demonstrate the ability of a quartz crystal microbalance to detect and to evaluate the association process occurring between molecules.

Journal Article↗

PubMed2006

Specific detection of glycans on a plasma membrane of living cells with atomic force microscopy.

Among the many alterations of cancer cells is the expression of different surface oligosaccharides. In this work, oligosaccharide expression in living cells (cancer and reference ones) was studied with atomic force microscopy by using lectins as probes. The unbinding force obtained for the same lectin type (concanavalin A or Sambucus nigra) suggested slightly dissimilar structures of binding sites of the same ligand type. For the lectin from Phaseolus vulgaris, a much larger unbinding force indicated a distinct structure of the binding site in cancer cells. The unbinding probability confirmed a higher content of both sialic acid and mannose-containing ligands in cancer and reference cells, respectively. These results demonstrate the potential of atomic force microscopy to directly probe the presence of molecules on a living cell surface, together with the quantitative description of their expression.

Cell Line, Tumor↗

PubMed2005

Friction force microscopy as an alternative method to probe molecular interactions.

Friction force microscopy was applied to study protein-carbohydrate interactions that are important in many cellular recognition processes. The expression and structure of carbohydrates can be investigated using lectins as molecular probes since they recognize different types of sugar molecules. Lectins (concanavalin A and lentil lectin, recognizing mannose-type carbohydrates) were attached to the probing tip and carboxypeptidase Y (possessing complementary carbohydrates) was immobilized on a modified glass surface using microcontact printing. The results obtained from friction force maps and dependencies on the loading rate (measured in a physiological buffer) were divided in two distinct groups. The first group of results obtained for lectin-protein complexes was assigned to molecular recognition events, whereas the other including all control measurements was attributed to nonspecific interaction. All results presented here indicate that friction force microscopy can be successfully employed to study recognition processes.

Journal Article↗

PubMed2005

Catalytically grown carbon nanotubes of small diameter have a high Young's modulus.

Experimental studies of carbon nanotubes (CNTs) obtained through different synthesis routes show considerable variability in their mechanical properties. The strongest CNTs obtained so far had a high Young's modulus of 1 TPa but could only be produced in gram scale quantities. The synthesis by catalytic chemical vapor deposition, a method that holds the greatest potential for large-scale production, gives CNTs with a high defect density. This leads to low Young's modulus values below 100 GPa for multiwall CNTs. Here we performed direct measurements of the mechanical properties of catalytically grown CNTs with only a few walls and find a Young's modulus of 1 TPa. This high value is confirmed for CNTs grown under two different growth conditions where the synthesis parameters such as the hydrocarbon source, catalyst material, and the synthesis temperature were varied. The results indicate that the observed difference in the Young's modulus for the catalytically grown CNTs with high and low numbers of walls is probably related to the growth mechanism of CNT.

Journal Article↗

PubMed2004

Hydrogel microspheres: influence of chemical composition on surface morphology, local elastic properties, and bulk mechanical characteristics.

Hydrogel microspheres, beads, and capsules of uniform size, differing in their chemical composition, have been prepared by electrostatic complex formation of sodium alginate with divalent cations and polycations. These have served as model spheres to study the influence of the chemical composition on both surface characteristics and bulk mechanical properties. Resistance to compression experiments yielding the compression work clearly identified differences as a function of the composition, with forces at maximal compression in the range of 34-455 mN. The suitability and informative value of atomic force microscopy have been confirmed for the case where surface characterization is performed in a liquid environment equivalent to physiological conditions. Surface imaging and mechanical response to indentation revealed different average surface roughness and Young's moduli for all hydrogel types ranging from 0.9 to 14.4 nm and from 0.4 to 440 kPa, respectively. The hydrogels exhibited pure elastic behavior. Despite a relatively high standard deviation, resulting from both surface and batch heterogeneity, nonoverlapping ranges of Young's moduli were reproducibly identified for the selected model spheres. The findings indicate the reliability of contact mode atomic force microscopy to quantify local surface properties, which may have an impact on the biocompatibility of alginate-based hydrogel materials of different composition and conditions of preparation. Moreover, it seems that local elastic properties and bulk mechanical characteristics are subject to analogous composition influences.

Journal Article↗

PubMedDate not supplied

Large fluctuations in the disassembly rate of microtubules revealed by atomic force microscopy.

Atomic force microscopy (AFM) in situ has been used to observe the cold disassembly dynamics of microtubules at a previously unrealised spatial resolution. Microtubules either electrostatically or covalently bound to aminosilane surfaces disassembled at room temperature under buffer solutions with no free tubulin present. This process was followed by taking sequential tapping-mode AFM images and measuring the change in the microtubule end position as a function of time, with an spatial accuracy down to +/-20nm and a temporal accuracy of +/-1s. As well as giving average disassembly rates on the order of 1-10 tubulin monomers per second, large fluctuations in the disassembly rate were revealed, indicating that the process is far from smooth and linear under these experimental conditions. The surface bound rates measured here are comparable to the rates for GMPCPP-tubulin microtubules free in solution, suggesting that inhibition of tubulin curvature through steric hindrance controls the average, relatively low disassembly rate. The large fluctuations in this rate are thought to be due to multiple pathways in the kinetics of disassembly with differing rate constants and/or stalling due to defects in the microtubule lattice. Microtubules that were covalently bound to the surface left behind the protofilaments covalently cross-linked to the aminosilane via glutaraldehyde during the disassembly process. Further work is needed to quantitatively assess the effects of surface binding on protofibril disassembly rates, reveal any differences in disassembly rates between the plus and minus ends and to enable assembly as well as disassembly to be imaged in the microscope fluid cell in real-time.

Aluminum Silicates↗