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Pyruvate Kinase: Atp:pyruvate 2-O-phosphotransferase. A phosphotransferase that catalyzes reversibly the phosphorylation of pyruvate to phosphoenolpyruvate in the presence of Atp. It has four isozymes (L, R, M1, and M2). Deficiency of the enzyme results in hemolytic anemia. EC 2.7.1.40.

What is Glycolysis?

JoVE 10737

Cells make energy by breaking down macromolecules. Cellular respiration is the biochemical process that converts “food energy” (from the chemical bonds of macromolecules) into chemical energy in the form of adenosine triphosphate (ATP). The first step of this tightly regulated and intricate process is glycolysis. The word glycolysis originates from Latin glyco (sugar) and lysis (breakdown). Glycolysis serves two main intracellular functions: generate ATP and intermediate metabolites to feed into other pathways. The glycolytic pathway converts one hexose (six-carbon carbohydrate such as glucose), into two triose molecules (three-carbon carbohydrate) such as pyruvate, and a net of two molecules of ATP (four produced, two consumed) and two molecules of nicotinamide adenine dinucleotide (NADH). Did you know that glycolysis was the first biochemical pathway discovered? In the mid-1800s, Louis Pasteur determined that microorganisms cause the breakdown of glucose in the absence of oxygen (fermentation). In 1897, Eduard Buchner found that fermentation reactions can still be carried out in cell-free yeast extracts, achieved by breaking open the cell and collecting the cytoplasm which contains the soluble molecules and organelles. Shortly thereafter in 1905, Arthur Harden and William Young discovered that the rate of fermentation decreases wit

 Core: Cellular Respiration

Energy-releasing Steps of Glycolysis

JoVE 10739

While the first phase of glycolysis consumes energy to convert glucose to glyceraldehyde 3-phosphate (G3P), the second phase produces energy. The energy is released over a sequence of reactions that turns G3P into pyruvate. The energy-releasing phase—steps 6-10 of glycolysis—occurs twice, once for each of the two 3-carbon sugars produced during steps 1-5.

The first energy-releasing step—considered the 6th step of glycolysis overall—consists of two concurrent events: oxidation and phosphorylation of G3P. The electron carrier NAD+ removes one hydrogen from G3P, oxidizing the 3-carbon sugar and converting (reducing) NAD+ to form NADH and H+. The released energy is used to phosphorylate G3P, turning it into 1,3-bisphosphoglycerate. In the next step, 1,3-bisphosphoglycerate converts ADP to ATP by donating a phosphate group, thereby becoming 3-phosphoglycerate. The 3-phosphoglycerate is then converted into an isomer, 2-phosphoglycerate. Subsequently, 2-phosphoglycerate loses a water molecule, becoming the unstable molecule 2-phosphoenolpyruvate, or PEP. PEP easily loses its phosphate group to ADP, converting it into a second ATP molecule and becoming pyruvate in the process. The energy-releasing phase releases two molecules of ATP and one molecule of NADH per converted sugar. Because

 Core: Cellular Respiration

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

1Department of Pediatric Oncology/Hematology, VU University Medical Center, 2Department of Hematology, VU University Medical Center, 3Department of Medical Oncology, VU University Medical Center, 4Department of Clinical Genetics, VU University Medical Center, 5Division of General and Transplant Surgery, Azienda Ospedaliera Universitaria Pisana, Universita’ di Pisa, 6Amsterdam Immunology and Rheumatology Center, VU University Medical Center, 7Princess Máxima Center for Pediatric Oncology, 8Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, 9Institute of Nanoscience and Nanotechnology, CNR-Nano

JoVE 54714

 Cancer Research

Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

1Centro Cardiologico Monzino IRCCS, 2Cardiovascular Tissue Bank of Milan, Centro Cardiologico Monzino IRCCS, 3Department of Clinical Sciences and Community Health, Cardiovascular Section, University of Milan, 4Department of Cardiovascular Disease, Development and Innovation Cardiac Surgery Unit, Centro Cardiologico Monzino IRCCS

JoVE 55762

 Biochemistry
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