Navigation Links
Preparative Nondenaturing Gel Electrophoresis to Purify NADP-Specific Glutamate Dehydrogenase From Chlorella, Rev B

Contributed by Philip W Miller, Waltraud I Dunn, and Robert R Schmidt, University of Florida, Dept. Microbiology and Cell Science, Gainesville, FL 32611 USA
Florida Agricultural Experiment Station Serial Number T-00256


Introduction
One of the research goals of this laboratory is to elucidate the pre- and posttranslational mechanisms that regulate both carbon and nitrogen metabolism in plants. Chlorella sorokiniana, a unicellular green alga, has been used extensively as a model system to study enzymes involved in higher plant metabolism. Research in this laboratory has shown that Chlorella sorokiniana possesses seven ammonium inducible, chloroplastic, NADP-specific glutamate dehydrogenase (NADP-GDH) isoenzymes that are regulated by both nitrogen and carbon metabolites (Prunkard et al. 1986). Biochemical, immunochemical, and physical characterization of purified GDH homohexamers revealed that the subunits (α, 55.5 kD; , 53 kD) composing these holoenzymes are nearly identical; however, kinetic characterization of the enzymes showed them to have strikingly different Km values, allosteric properties, and turnover rates (Bascomb and Schmidt 1987). Recent molecular genetic analysis indicates that minor modifications in the primary protein sequences may account for these observed properties. In order to further elucidate the regulation of these isoenzymes, it was necessary to purify to homogeneity a large quantity of active enzyme for polyclonal/monoclonal antibody production and additional protein analysis.

We report here the use of the Model 491 prep cell for the purification of native NADP-GDH protein by nondenaturing PAGE with <2% loss
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
2. Detection of Platinum Species in Plant Material by Preparative Isotachophoresis Using the Model 491 Prep Cell, Rev A
3. Preparative Nondenaturing Gel Electrophoresis Used in the Purification of an Esterase Involved in Insecticide Resistance, Rev C
4. Preparative SDS-PAGE Electrophoresis of a Recombinant Epstein-Barr Virus Encoded Protein and Its Application in Serodiagnostic Test Systems, Rev C
5. Purification and Characterization of beta-Lactoglobulin Genetic Variants A and B Using Preparative Electrophoresis and Isoelectric Focusing
6. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
7. Semi Preparative Purification of Proteins
8. A Versatile Power Supply for All Electrophoresis Applications
9. A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis
10. Continuous-Elution Electrophoresis for Purification of the Baculovirus-Expressed Coronavirus Structural Proteins, Rev A
11. Detection of K-ras Point Mutations in the Pancreas by Constant Denaturing Gel Electrophoresis Using the DCode System
Post Your Comments:
(Date:7/25/2008)...esident Expected To Regain Hearing in Both Ears fo...0 Years, DENVER, July 25 /- Cochlear...ies, today announced a landmark milestone when,Dr....ter performs the,120,000th cochlear implant device...Susan Grossman, will undergo surgery to receive bi...
(Date:7/25/2008)... A recent survey on creativity, spons...n and conducted by TNS Teleseker, revealed that a ...d a willingness to use special methods that could ... in this state for as long as desired." , ...ly 26, 2008 -- A recent survey on creativity, spon...
(Date:7/24/2008)...O, July 24, 2008 /- NuVasiv... focused on developing products for,minimally disr...d today,that it has completed the previously annou...s from Osiris Therapeutics, Inc. (Nasdaq: OSIR ) ...-based contingent payments not to,exceed $50 milli...
(Date:7/24/2008)...s hospital physicians, pulmonary and critical care...bout overuse of antibiotics, DURHAM, N.C., July 2...o,surveys, fielded during conferences of the Ameri...ine, that show how hospitalists, pulmonary and,cri...us in their view that,new methods and technologies...
Breaking Biology Technology:Cochlear Announces Milestone in Hearing Restoration With 120,000 Nucleus(R) Cochlear Implants 2Cochlear Announces Milestone in Hearing Restoration With 120,000 Nucleus(R) Cochlear Implants 3Cochlear Announces Milestone in Hearing Restoration With 120,000 Nucleus(R) Cochlear Implants 4Project Mind Survey of Israeli Ph.D. Scientists Favors a New Standard of Creativity in Science 2Project Mind Survey of Israeli Ph.D. Scientists Favors a New Standard of Creativity in Science 3NuVasive Completes Acquisition of Osteocel Biologics Business 2NuVasive Completes Acquisition of Osteocel Biologics Business 3NuVasive Completes Acquisition of Osteocel Biologics Business 4NuVasive Completes Acquisition of Osteocel Biologics Business 5U.S. Clinicians Indicate a Strong Need for New Tools to Combat Sepsis 2U.S. Clinicians Indicate a Strong Need for New Tools to Combat Sepsis 3
... Milwaukee investor group that has weathered a cou... leadership of tech entrepreneur Pehr Anderson. , ...s with high net worth, allows investors to pool st...ve investments. It began in 2000 with 13 investors...vestors made fewer deals over the last two years. ...
...ails for video) In each of these video clips, liv... pair of electrodes where they are trapped by mild...Taking a new approach to the painstaking assembly ...an hair scientists at the University of Wisconsin...ells to make tiny bio-electronic circuits. , ,The ...
...promising research breakthrough on Alzheimer,s dis...unded Mithridion Inc. in hopes of commercializing ...low researcher Thor Stein led a team that began ab...y don,t mouse neurons die when slammed by a protei...onset Alzheimer,s patients? From there, Johnsons t...
...rked on by state legislators would require organiz...n residents to notify those residents if their per...rty. , ,LRB-2214, recently introduced by state Sen...zgerald, aims to protect Wisconsin residents from ...ast month by giant California data aggregator Cho...
Other Biology Technology:Boston entrepreneur to enliven Milwaukee angel group 2New nanotech approach could enable fast anthrax detection 2Madison-based Mithridion seeks to capitalize on Alzheimer's breakthrough 2Madison-based Mithridion seeks to capitalize on Alzheimer's breakthrough 3Madison-based Mithridion seeks to capitalize on Alzheimer's breakthrough 4Bill would require data keepers to notify consumers of info leaks 2
(Date:7/24/2008)... by Massachusetts General Hospital (MGH) investiga... research and medicine a robust method for genera...DNA sequences for inactivation or repair. In the...s describe an efficient, publicly available method...s), which can be used to induce specific genomic m...
(Date:7/24/2008)...researchers from Columbia University,s Mailman Sch...llenging issues in infant health and preventing mo... In these settings, HIV-infected mothers had been ...s, they should exclusively breast-feed, followed b...But according to the study conducted in Lusaka, Za...
(Date:7/24/2008)...searchers have found a molecular link between circ... The discovery reveals new possibilities for the t...seases. , Paolo Sassone-Corsi, Distinguished Pro...es have identified that an essential protein calle...s, works in balance with another protein called SI...
(Date:7/24/2008)...A fire is currently burning through a study area w...ut 2 years ago. Managers used data and analysis fr...dy in the planning, analysis, and implementation o...ow active., The Gotchen LSR, lies on the east sl... about 15,000 acres of the Mount Adams Ranger Dist...
Breaking Biology News(10 mins):Consortium develops new method enabling routine targeted gene modification 2Early cessation of breastfeeding by HIV+ women in poor countries and child survival 2Early cessation of breastfeeding by HIV+ women in poor countries and child survival 3Circadian rhythm-metabolism link discovered 2Landscape study may offer solutions for fire managers 2Muscle Weakness Found in Some Autistic Children 16649 1Muscle Weakness Found in Some Autistic Children 16649 2Muscle Weakness Found in Some Autistic Children 16649 3Extrel CMS Announces ATEX Approval for Process Mass Spectrometer 16647 1Extrel CMS Announces ATEX Approval for Process Mass Spectrometer 16647 2Diabetic Food Shopping That Wont Break a Budget 16645 1Diabetic Food Shopping That Wont Break a Budget 16645 2Spring and Pollen Are in the Air 16641 1Spring and Pollen Are in the Air 16641 2
...Dana-Farber Cancer Institute and collaborators hav...tiple myeloma, a deadly blood cancer, that have di...ively with drugs specifically targeted to those su...orithm designed to recognize human faces plucked t...scape of many DNA alterations in the myeloma genom...
...e determined the detailed structure of an essentia...tributor to the vast majority of human cancers. Un...led to a better understanding of how it acts to im... broadly effective cancer drugs. , Until now, the ...enzyme has hindered progress in developing agents ...
...e determined the detailed structure of an essentia...tributor to the vast majority of human cancers. Un...led to a better understanding of how it acts to im... broadly effective cancer drugs. , Until now, the ...enzyme has hindered progress in developing agents ...
...more than three years, New York City,s Office of C... of the 9/11 World Trade Center attack victims, th...tergovernmental and family cooperation and the per...ing to a recent book by Dr. Robert C. Shaler, prof...rector of the forensics science major program at P...
Other Biology News:Scientists discover new genetic subtypes of common blood cancer 2Scientists discover new genetic subtypes of common blood cancer 3Report lists top 20 most-vulnerable African carnivores 2Report lists top 20 most-vulnerable African carnivores 3Report lists top 20 most-vulnerable African carnivores 4Groovy protein essential for promoting cancer development 2Groovy protein essential for promoting cancer development 3Groovy protein essential for promoting cancer development 4World Trade Center identifications pushed forensic DNA technology 2World Trade Center identifications pushed forensic DNA technology 3
...n or in a gel piece, we can identify it using MALD...sted to produce a mixture of peptides. The molecul...y MALDI-TOF, and the amino acid sequences of the t...rmined by MALDI-TOF/TOF. The masses and sequences ...
...e cytopathic effect inhibition assays, also known ...; tissue culture supernatents; serum from PK studi...e unit is defined as the quantity of interferon re...ect of viral infection by 50%. All standards use...
supplied with 10x reaction buffer
HyPro 20 system for slide hybridization. 20 slide capacity. Active vibration system, slide rack and humidity chamber. Ideal for microarray and in situ hybridization.
Biology Products: