The Uab Research Foundation

Other

The University of Alabama

Colorado State University Research Foundation

Mississippi State University

The United States of America As Represented by the Administrator of the

Virtual Drug Development, Inc.

Lawrence James DeLucas

Wayne J Brouillette

Christie G Brouillette

Terry L Bray

Charles Sherman Henry

Donald D Muccio

Sadanandan Velu

Laurent Schmitt

Wilbur W Wilson

David Johnson

Mark J Jedrzejas

Yancho Devedjiev

Jose Gabriel Garcia

Walter Cristofoli

Lisa Nagy

Daniel C Carter

Arnon Chait

Michael Hugh Harrington

Ming Luo

Robert S Snyder

Alexander I Belgovskiy

Charles E Daitch

Bouvard Hosticka

Thomas E Gester

Charles E Bugg

David T Hamrick

Vaughn H Yost

Brad Stoops

Blair J Herren

L Roger Mason, Jr

Pamela N Norris

Yong-Chul Kim

Fred L Suddath

Liyuan Mou

John E Volanakis

R Stephen Porter

Sadanadan E Velu

Jack S Brenizer, Jr

Larry Cosenza

Larry J Kim

The present disclosure provides a method to allow a user to pre-screen numerous crystallization conditions in the crystallization space to identify those conditions with the highest probability of yielding crystals and high quality diffracting crystals. In one embodiment, the dilute solution thermodynamic virial coefficient, termed B, is used to aid in the determination crystallization conditions that increase the probability of producing crystals for the crystallant of interest. The present disclosure also provide methods for predicting solution conditions that generate beneficial solubility and/or stability conditions for a polypeptide of interest using the B parameter. Devices for use in the described methods are also described.

Method for determining crystallization parameters and apparatus for use with the same

A method of screening protein crystal growth conditions with picogram to microgram amounts of protein in picoliter or nanoliter volumes is provided. A preferred method comprises a microarray with a plurality of microchambers in the microarray. A protein solution is placed into the micro-chambers by an automated dispensing mechanism. The protein crystal growth conditions of each of the micro-chambers is adjusted so that the protein crystal growth conditions in at least two of the micro-chambers differs. Crystallization of the protein solution in the micro-chambers is effected. For example, crystallization can be effected by a precipitate solution and/or placing an oil barrier over the protein solution. Protein crystal growth in the micro-chambers is then observed.

Method for screening crystallization conditions in solution crystal growth

An improved method of screening crystal growth conditions is provided wherein molecules are crystallized from solutions containing dyes. These dyes are selectively incorporated or associated with crystals of particular character thereby rendering crystals of particular character colored and improving detection of the dyed crystals. A preferred method involves use of dyes in protein solutions overlayed by oil. Use of oil allows the use of small volumes of solution and facilitates the screening of large numbers of crystallization conditions in arrays using automated devices that dispense appropriate solutions to generate crystallization trials, overlay crystallization trials with an oil, provide appropriate conditions conducive to crystallization and enhance detection of dyed (colored) or undyed (uncolored) crystals that result.

Use of dye to distinguish salt and protein crystals under microcrystallization conditions

A method of screening protein crystal growth conditions with picogram to microgram amounts of protein in picoliter or nanoliter volumes is provided. A preferred method comprises a microarray with a plurality of micro-chambers in the microarray. A protein solution is placed into the micro-chambers by an automated dispensing mechanism. The protein crystal growth conditions of each of the micro-chambers is adjusted so that the protein crystal growth conditions in at least two of the micro-chambers differs. Crystallization of the protein solution in the micro-chambers is effected. Protein crystal growth in the micro-chambers is then observed.

A method of screening solution conditions suitable for the crystallization of macromolecules with a picogram to a microgram of protein using picoliter or nanoliter volumes is provided. A preferred method comprises preparing a plurality of recipe solutions in milliliter volumes, aspirating protein and recipe solutions, respectively, from a plurality of wells, and dispensing the protein and recipe solutions into a plurality of wells, covering the combined protein and recipe solutions with oil, and maintaining the solution wells until crystallization or precipitation occur therein.

Method for preparation of microarrays for screening of crystal growth conditions

A method of screening protein crystal growth conditions on a nano or meso scale comprises providing a micro-electromechanical chip with a plurality of micro-chambers in the micro-electromechanical chip. A protein solution is placed into the micro-chambers by an automated jet type dispensing means with nano or meso scale precision. The protein crystal growth conditions of each of the micro-chambers is adjusted so that the protein crystal growth conditions of each of the micro-chambers differs. Crystallization of the protein solution in the micro-chambers is effected. Protein crystal growth in the micro-chambers is then observed.

Disclosed are compounds that inhibit the microbial NAD synthetase enzyme. For example, disclosed are compounds of the formula Ar&#x2014;X&#x2014;Ar&#x2014;Y&#x2014;L&#x2014;Z&#x2014;Q, wherein Q is QAror ArQ; Ar, Ar, and Arare independently aryl or heteroaryl, optionally substituted with one or more substituents; X, Y, and Z are independently selected from the group consisting of a covalent bond or groups containing one or more of C, H, N, O, S atoms; L is a linker and Qis an alkylenyl, alkylenyl carbonyloxy alkyl, or alkylenyl carbonylamino alkyl group, optionally having a substituent; a covalent bond; a group containing amidine or guanidine function wherein the amidine or guanidine may be optionally N-substituted with an alkyl; or a zwitterion; or a pharmaceutically acceptable salt thereof. Also disclosed are methods which involve the use of the compounds of the present invention, for example, in treating or preventing a microbial infection in a mammal or plant, killing a prokaryote or decreasing prokaryotic growth, disinfecting a material or environment contaminated by a microbe, increasing food animal production, controlling harm to plants by a pest or insect, and combating agroterrorism. Examples of microbes affected by the compounds of the present invention are bacteria and fungi.

NAD synthetase inhibitors and uses thereof

The present invention provides methods of synthesizing and screening inhibitors of bacterial NAD synthetase enzyme, compounds thereof, and methods of treating bacterial and microbial infections with inhibitors of bacterial NAD synthetase enzyme.

Inhibitors of bacterial NAD synthetase

The present invention provides methods of treating or preventing fungal infections in a host comprising administering a treatment effective or treatment effective amount of a yeast NAD synthetase inhibitor compound. The invention further provides a method of killing yeast comprising administering a yeast NAD synthetase compound that selectively binds to catalytic sites in yeast whereby the yeast is killed.

Methods of treating fungal infections with inhibitors of NAD synthetase enzyme

Smart aerogel, an aerogel material doped with special bio-affinity compounds to providing means of unique collection, detection and identification of bioaerosols, including bacteria, viruses, toxins, and other bioaerosols. Aerogels, extremely low density and highly porous materials with a complex pore structure, are used as an intelligent detection material by incorporating specific bioaffinity pharmaceuticals directly into the matrix. The complex pore structure contains micropores, mesopores, and macropores in an open pore structure. The opening pore structure of the aerogel is used to create docking sites by linking high affinity pharmaceuticals that specifically bind only to certain bioaerosols. The high internal surface area of the aerogel and the extremely low density provides abundant receptor sites per unit mesopore for a high bioaerosol-receptor interaction, yet in a manner which will reduce possible damage and destruction to the bioaerosols captured.

Smart aerogel

Crystal growth can be initiated and controlled by dynamically controlled vapor diffusion or temperature change. In one aspect, the present invention uses a precisely controlled vapor diffusion approach to monitor and control protein crystal growth. The system utilizes a humidity sensor and various interfaces under computer control to effect virtually any evaporation rate from a number of different growth solutions simultaneously by means of an evaporative gas flow. A static laser light scattering sensor can be used to detect aggregation events and trigger a change in the evaporation rate for a growth solution. A control/follower configuration can be used to actively monitor one chamber and accurately control replicate chambers relative to the control chamber. In a second aspect, the invention exploits the varying solubility of proteins versus temperature to control the growth of protein crystals. This system contains miniature thermoelectric devices under microcomputer control that change temperature as needed to grow crystals of a given protein. Complex temperature ramps are possible using this approach. A static laser light scattering probe also can be used in this system as a non-invasive probe for detection of aggregation events. The automated dynamic control system provides systematic and predictable responses with regard to crystal size. These systems can be used for microgravity crystallization projects, for example in a space shuttle, and for crystallization work under terrestial conditions. The present invention is particularly useful for macromolecular crystallization, e.g. for proteins, polypeptides, nucleic acids, viruses and virus particles.

Dynamically controlled crystal growth system

Crystals of human complement factor D that are triclinic are disclosed. They are made by a method of crystallization known as the 'hanging drop' technique. The technique uses 1.0-1400 mM NaCl and polyethylene glycol maintained at a pH of 5.2-7.0, more preferably 5.3-6.1.

Crystals of human complement factor D that are triclinic

A macromolecular crystal growing system especially designed for growing crystals in the low gravity of space as well as the gravity of earth includes at least one tray assembly, a carrier assembly which receives the tray, and a refrigeration-incubation module in which the carrier assembly is received. The tray assembly includes a plurality of sealed chambers with a plastic syringe and a plug means for the double tip of the syringe provided therein. Ganging mechanisms operate the syringes and plugs simultaneously in a precise and smooth operation. Preferably, the tray assemblies are mounted on ball bearing slides for smooth operation in inserting and removing the tray assemblies into the carrier assembly. The plugging mechanism also includes a loading control mechanism. A mechanism for leaving a syringe unplugged is also provided.

Macromolecular crystal growing system

Growing community of inventors

Birmingham, AL, United States of America

Lawrence James DeLucas