Biomedical subjects

J Mou

Publications and source records attributed to J Mou.

12 recordsLinked to original sources

PubMed1999

[Determination of pentachlorophenol residues in textiles and leather and leather products by gas chromatography/mass spectrometry].

A method for determination of pentachlorophenol residues in textiles and leather and leather products by gas chromatography/mass spectrometry was developed. The pentachlorophenol residues in samples was acidified with sulphuric acid solution (6 mol/L) and extracted with n-hexane. After the n-hexane layer was washed with sodium sulphate solution (20 g/L)until it was clear, determination was made by means of a gas chromatograph equipped with MSD, using external standard method. A DB-17 fused silica capillary column (30 m x 0.25 mm i.d.; 0.25 micron film thickness) was employed in the analysis, The column temperature program included a 2 min isothermal period at 50 degrees C, temperature increased at a rate of 30 degrees C/min to 220 degrees C, and hold for 1 min; temperature increased of at a rate 6 degrees C/min to 260 degrees C, and hold for 1 min. Mass spectra were obtained by electron impact at 70 eV and the determination was made by means of SIM mode, selected monitoring ion (m/z) was 264 amu. The detection limit was 0.02 mg/kg. The average recovery was from 86.7% to 93.1% and CV (n = 10) is from 4.1% to 5.9%. This method was already used to determine the mass concentration of pentachlorophenol residues in raw cotton, towel, shirt, leather gloves, leather watchguard. The method is simple, rapid and accurate.

Environmental Pollutants↗

PubMed1996

Gramicidin A aggregation in supported gel state phosphatidylcholine bilayers.

Using an atomic force microscope, supported bilayers of saturated phosphatidylcholine (in the gel state) containing various amounts of gramicidin A (gA) were imaged in aqueous solutions and at room temperature. gA clusters were directly observed for the first time under these conditions. It was found that, at a lower gA concentration, gA aggregated into domains, composed of small clusters along with a considerable amount of lipids. This basic aggregation unit, most likely a hexamer, remained the same for acyl chain lengths from 14 to 18 carbons. These small clusters were observed to form elongated aggregates (line type) but never into extended pure gA domains. When gA concentrations were increased, for bilayers with 16 carbons or less, gA aggregated into larger domains but the basic unit remained separated by lipid molecules. At about 5 mol % gA, a percolation-like transition occurred at which the line type aggregates were connected to each other. However, for bilayers with more than 16 carbons, multiple lamellar structures were formed at higher gA fractions and the top layer had a ripple-like surface morphology. The molecular mechanism for the formation of these peculiar structures remains to be elucidated.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1996

High resolution surface structure of E. coli GroES oligomer by atomic force microscopy.

Using atomic force microscopy (AFM) in aqueous solution, we show that the surface structure of the oligomeric GroES can be obtained up to 10 angstroms resolution. The seven subunits of the heptamer were well resolved without image averaging. The overall dimension of the GroES heptamer was 8.4 +/- 0.4 nm in diameter and 3.0 +/- 0.3 nm high. However, the AFM images further suggest that there is a central protrusion of 0.8 +/- 0.2 nm high and 4.5 +/- 0.4 nm in diameter on one side of GroES which displays a profound seven-fold symmetry. It was found that GroEL could not bind to the adsorbed GroES in the presence of AMP-PNP and Mg2+, suggesting that the side of GroES with the central protrusion faces away from the GroEL lumen, because only one side of GroES was observed under these conditions. Based on the results from both electron and atomic force microscopy, a surface model for the GroES is proposed.

Bacterial Proteins↗

PubMed1996

Chaperonins GroEL and GroES: views from atomic force microscopy.

The Escherichia coli chaperonins, GroEL and GroES, as well as their complexes in the presence of a nonhydrolyzable nucleotide AMP-PNP, have been imaged with the atomic force microscope (AFM). We demonstrate that both GroEL and GroES that have been adsorbed to a mica surface can be resolved directly by the AFM in aqueous solution at room temperature. However, with glutaraldehyde fixation of already adsorbed molecules, the resolution of both GroEL and GroES was further improved, as all seven subunits were well resolved without any image processing. We also found that chemical fixation was necessary for the contact mode AFM to image GroEL/ES complexes, and in the AFM images. GroEL with GroES bound can be clearly distinguished from those without. The GroEL/ES complex was about 5 nm higher than GroEL alone, indicating a 2 nm upward movement of the apical domains of GroEL. Using a slightly larger probe force, unfixed GroEL could be dissected: the upper heptamer was removed to expose the contact surface of the two heptamers. These results clearly demonstrate the usefulness of cross-linking agents for the determination of molecular structures with the AFM. They also pave the way for using the AFM to study the structural basis for the function of GroE system and other molecular chaperones.

Adenylyl Imidodiphosphate↗

PubMed1996

The effect of deformation on the lateral resolution of atomic force microscopy.

A computer model based on the elastic properties of rubber is introduced for the evaluation of the lateral resolution in atomic force microscopy of deformable specimens. The computational results show that, if the full width at half-height can be defined as the lateral resolution, it is continuously improved at greater probe forces, at the expense of a reduced molecular height, In fact, even for a probe that is bigger than the molecule, the real size of the molecule can be 'recovered' at about 25% compression. This result demonstrates that for a better lateral resolution, a greater probe force can be beneficial, provided that the molecule is not moved or damaged and the response remains elastic. Measurements on isolated low-density lipoproteins (LDL) show that with 26% vertical compression, the lateral size measured in atomic force microscopy is only 72% of the value predicted by a simple convolution, and is only slightly larger (approximately 13%) than the known size of LDL. Therefore, the results on LDL provide a direct support for the conclusions of the computational model.

Computer Simulation↗

PubMed1995

High-resolution atomic-force microscopy of DNA: the pitch of the double helix.

Using a cationic lipid bilayer, we show that DNA can be reliably adsorbed to the bilayer surface for atomic force microscopy (AFM) in aqueous buffers at high resolution. The measured width of the dsDNA is close to 2 nm, and a periodic modulation on dsDNA is reproducibly detected by the AFM. The measured period is 3.4 +/- 0.4 nm, in excellent agreement with the known pitch of the double helix. The right-handedness of the double helix is directly discernible in high resolution AFM images. Thus, this approach can be readily applied to the study of DNA-protein interactions, as well as sequence mapping at high resolution.

DNA↗

PubMed1995

Cryo atomic force microscopy: a new approach for biological imaging at high resolution.

A low-temperature atomic force microscope (cryo-AFM), operated in liquid nitrogen vapor, has been constructed for biological applications. The system provides an adjustable imaging temperature from 77 to 220 K with atomic resolution achieved on crystalline specimens. Imaging with NaCl microcrystals demonstrates that the system is free from surface contamination. Below 100 K, several biological specimens, including immunoglobulins and DNA as well as red blood cell ghosts, were imaged at high spatial resolution. Measurements on individual macromolecules showed that the mechanical strength is significantly greater at cryogenic temperatures with an estimated Young's modulus 1000-10,000 times that of a hydrated protein at room temperature, providing a solid basis for future improvements and applications of cryo-AFM in structural biology.

DNA↗

PubMed1995

Atomic force microscopy of cholera toxin B-oligomers bound to bilayers of biologically relevant lipids.

Cholera toxin B-oligomer was imaged by atomic force microscopy (AFM) on biologically relevant model membranes, such as 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine and egg yolk phosphatidylcholine at room temperature in solution at a resolution in the range of 1 to 2 nm. In addition, two-dimensional arrays were grown directly on these model membranes without any special treatment, and were also imaged by AFM. These results demonstrate the ability of AFM for imaging membrane proteins at high resolution without the need of chemical cross-linking, either within the membrane or to the substratum.

Cholera Toxin↗

PubMed1994

Alcohol induces interdigitated domains in unilamellar phosphatidylcholine bilayers.

Using the atomic force microscope (AFM) in situ, we have demonstrated that acyl chain interdigitation can be induced reversibly by alcohol in supported unilamellar phospholipid bilayers. At alcohol concentrations considerably lower than the critical values determined by other experimental techniques, it was found that interdigitated domains can be induced and these domains were stable over a long period of time. The mechanism of such domain formation remains to be elucidated. This work also serves as an example to illustrate the uniqueness of the AFM as a powerful tool in the study of membrane structure and conformation under physiological conditions at high spatial resolution.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1994

Tris(hydroxymethyl)aminomethane (C4H11NO3) induced a ripple phase in supported unilamellar phospholipid bilayers.

A commonly used buffer compound, tris(hydroxymethyl)aminomethane (C4H11NO3), was found to induce a ripple phase in supported unilamellar phospholipid bilayers at room temperature. The ripple structure showed various types of domains that could extend to several micrometers in length with many well-defined bendings of either 120 degrees or 60 degrees. Two different periodic ripples were found to coexist in 1,2-dipentadecanoyl-sn-glycero-3-phosphatidylcholine bilayers. Some intermediate states during the transition to the ripple phase were also observed.

Chemical Phenomena↗

PubMed1994

Molecular resolution atomic force microscopy of soluble proteins in solution.

We introduce a simple specimen preparatory method for atomic force microscopy of soluble proteins in aqueous solutions. It is demonstrated that the mica surface is suitable for direct adsorption of macromolecules that are sufficiently stable to withstand the disturbance of the probe for reproducible imaging at high resolution. It is also shown that the main problem impeding successful imaging is the excessive adsorption of macromolecules, as loosely bound macromolecules readily stick to the tip and produce various imaging artifacts.

Animals↗

PubMed1994

Structure and stability of pertussis toxin studied by in situ atomic force microscopy.

Pertussis toxin, both complete and the B-oligomer, were imaged by atomic force microscopy (AFM), using specimens prepared by simple surface adsorption on mica without further manipulation. The spatial arrangement of the subunits of the B-oligomer was clearly resolved, representing the first protein quaternary structure obtained by AFM in situ. The results suggest that the B-oligomer is a flat pentamer with the two large subunits located next to each other, and the catalytic A-subunit situated at the center above. We found that the B-pentamer was structurally stable for temperatures up to 60 degrees C and within the pH range of 4.5-9.5. It is also demonstrated that the AFM was capable of resolving features down to 0.5 nm on the B-oligomers, indicating its great potential for structural determination.

Hot Temperature↗