Purification of homogeneous rat phosphofructokinase isozymes with high specific activities

Biochemical and Biophysical Research Communications
T P KastenG A Dunaway

Abstract

The purification of rat muscle and liver phosphofructokinase (PFK) isozymes has been greatly facilitated by column chromatographic separation on immobilized Cibacron Blue F3GA. The homogeneous liver PFK isozyme exhibited a specific activity of greater than 200 units per mg of protein which is nearly two-fold greater than has been previously reported for this isozyme. The yields for this isozyme exceeded 40% of the original activity and the molecular weight of its subunit was about 85,000 as determined by SDS-polyacrylamide gel electrophoresis. The muscle PFK isozyme's specific activity was approximately 265 units/mg of protein which also is about twice the greatest specific activity previously reported. The overall yield for muscle PFK exceeded 50% of the original activity, and the molecular weight of its subunit was approximately 82,000. Using each homogeneous isozyme, antibodies were produced in rabbits; and the immunoglobin-G (IgG) fraction from the sera of these rabbits was highly specific for the PFK isozyme used as an antigen.

References

Apr 26, 1989·Biochimica Et Biophysica Acta·E C HeesbeenG E Staal
Jan 1, 1989·The International Journal of Biochemistry·E C HeesbeenG E Staal
Aug 1, 1992·The International Journal of Biochemistry·E Fukushima, H Sugiya
Sep 1, 1986·Mechanisms of Ageing and Development·G A DunawayJ A Chesky
Jan 1, 1995·Neurochemistry International·Marina Bigl, Klaus Eschrich
Jan 1, 1989·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·T KoyamaE Iizuka
Jan 1, 1997·Free Radical Biology & Medicine·I MiwaM Sugiura
Apr 13, 2000·The International Journal of Biochemistry & Cell Biology·M SawadaS Furuyama
Dec 1, 1990·Differentiation; Research in Biological Diversity·H E van ErpG E Staal
Dec 15, 1994·European Journal of Biochemistry·O H Martínez-CostaJ J Aragón
Jun 5, 2013·Biological Chemistry·Torsten SchönebergNorbert Sträter

Citations

Related Concepts

August Rats
Cross Reactions
Phosphoglycerate Dehydrogenase Activity
Phosphofructokinase
Pfkp
Cibacron Blue F 3GA
Antigens
Muscle
Purification Aspects
Pfkl

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Synapse Loss as Therapeutic Target in MS

As we age, the number of synapses present in the human brain starts to decline, but in neurodegenerative diseases this occurs at an accelerated rate. In MS, it has been shown that there is a reduction in synaptic density, which presents a potential target for treatment. Here is the latest research on synapse loss as a therapeutic target in MS.

Artificial Intelligence in Cardiac Imaging

Artificial intelligence (ai) techniques are increasingly applied to cardiovascular (cv) medicine in cardiac imaging analysis. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

Social Learning

Social learning involves learning new behaviors through observation, imitation and modeling. Follow this feed to stay up to date on the latest research.

Cell Atlas of the Human Eye

Constructing a cell atlas of the human eye will require transcriptomic and histologic analysis over the lifespan. This understanding will aid in the study of development and disease. Find the latest research pertaining to the Cell Atlas of the Human Eye here.

Single Cell Chromatin Profiling

Techniques like ATAC-seq and CUT&Tag have the potential to allow single cell profiling of chromatin accessibility, histones, and TFs. This will provide novel insight into cellular heterogeneity and cell states. Discover the latest research on single cell chromatin profiling here.

Genetic Screens in iPSC-derived Brain Cells

Genetic screening is a critical tool that can be employed to define and understand gene function and interaction. This feed focuses on genetic screens conducted using induced pluripotent stem cell (iPSC)-derived brain cells.

© 2020 Meta ULC. All rights reserved