Investigation of the Transient State of an Allosteric Model with a Two-Stage Substrate Binding

Our previous results indicate that two types of fibronectin-cytoskeletal associations may be formed at the fibroblast surface: dorsal matrixbinding fibronexuses generated in high serum (5% FBS) cultures, and ventral substrate-adhering units formed in low serum (0.3% FBS) cultures. The substrate-adhering fibronexus consists of at least vinculin (VN) and actin in its cytoplasmic leg, and fibronectin (FN) as one of its major extracellular components. This substrate-adhesion complex is localized in focal contacts, the sites of closest substratum approach visualized with interference reflection microscopy, which appear to be the major points of cell-tosubstrate adhesion. In fibroblasts, the latter substrate-binding complex is characteristic of cultures that are arrested at the G1 phase of the cell cycle due to the low serum concentration in their medium. These arrested fibroblasts are very well spread, flattened, and immobile.

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Analyzing reactions of single mechanoenzyme molecules by light microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122538 ◽

1992 ◽

Vol 50 (1) ◽

pp. 426-427

Author(s):

Jeff Gelles

Keyword(s):

Image Processing ◽

Reaction Mechanisms ◽

Transient State ◽

Nanometer Scale ◽

Reaction Intermediates ◽

Enzyme Molecule ◽

Directed Movement ◽

Enzyme Molecules ◽

Individual Enzyme ◽

Single Reaction

Mechanoenzymes are enzymes which use a chemical reaction to power directed movement along biological polymer. Such enzymes include the cytoskeletal motors (e.g., myosins, dyneins, and kinesins) as well as nucleic acid polymerases and helicases. A single catalytic turnover of a mechanoenzyme moves the enzyme molecule along the polymer a distance on the order of 10−9 m We have developed light microscope and digital image processing methods to detect and measure nanometer-scale motions driven by single mechanoenzyme molecules. These techniques enable one to monitor the occurrence of single reaction steps and to measure the lifetimes of reaction intermediates in individual enzyme molecules. This information can be used to elucidate reaction mechanisms and determine microscopic rate constants. Such an approach circumvents difficulties encountered in the use of traditional transient-state kinetics techniques to examine mechanoenzyme reaction mechanisms.

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Two-stage self-seeding method for preparation of single crystals of poly(phenylene sulfide)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100153816 ◽

1989 ◽

Vol 47 ◽

pp. 368-369

Author(s):

Sengshiu Chung ◽

Peggy Cebe

Keyword(s):

Single Crystals ◽

High Performance ◽

Dilute Solution ◽

Two Stage ◽

Annealing Behavior ◽

High Performance Polymers ◽

Seeding Method ◽

Phenylene Sulfide

We are studying the crystallization and annealing behavior of high performance polymers, like poly(p-pheny1ene sulfide) PPS, and poly-(etheretherketone), PEEK. Our purpose is to determine whether PPS, which is similar in many ways to PEEK, undergoes reorganization during annealing. In an effort to address the issue of reorganization, we are studying solution grown single crystals of PPS as model materials.Observation of solution grown PPS crystals has been reported. Even from dilute solution, embrionic spherulites and aggregates were formed. We observe that these morphologies result when solutions containing uncrystallized polymer are cooled. To obtain samples of uniform single crystals, we have used two-stage self seeding and solution replacement techniques.

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