PMID: 9428658Jan 15, 1998Paper

Expression in insect cells and characterization of the 110 kDa anchoring subunit of myosin light chain phosphatase

Biochimica Et Biophysica Acta
Y ZhangT Tao

Abstract

The major myosin light chain phosphatase is composed of three subunits with apparent molecular masses of 130, 38 and 20 kDa, corresponding to the myosin-binding, catalytic and a regulatory subunit of unknown function, respectively. In this work, we have amplified the cDNA coding for each of the three subunits by the polymerase chain reaction, and expressed the 130 kDa subunit in insect cells using the baculovirus expression system. Limited chymotrypsin digestion show that the folding of the expressed protein is similar to that in the native holoenzyme. N-Terminal sequencing reveals that our recombinant protein is authentic. Mass spectrometry shows that the expressed protein is full length. The recombinant protein is capable of binding myosin based on the ELISA assay and myosin affinity chromatography. Finally, rotary shadowing electron microscopy reveals an elongated structure with three globular domains connected by flexible strands. These results pave the way for future biochemical, structural and site-directed mutagenesis studies on the myosin light chain phosphatase. We also found that the cDNA of the 20 kDa subunit may code for a smaller protein with a molecular mass of 18.5 kDa.

References

Jan 1, 1976·Journal of Biochemistry·S Ebashi
May 1, 1990·Journal of Structural Biology·K Mabuchi
Jan 1, 1986·Journal of Molecular and Cellular Cardiology·P J SilverJ T Stull
Jan 1, 1980·Methods in Enzymology·E Engvall
May 14, 1993·Biochemical and Biophysical Research Communications·H ShimaM Nagao
Nov 17, 1994·Nature·A P Somlyo, A V Somlyo
Apr 15, 1994·Biochemical and Biophysical Research Communications·S OkuboD J Hartshorne
Nov 22, 1993·FEBS Letters·P T Szymanski, T Tao
May 1, 1993·Trends in Biochemical Sciences·M J Hubbard, P Cohen
May 21, 1996·Biochemistry·K IchikawaD J Hartshorne
Aug 14, 1996·Biochemical and Biophysical Research Communications·M Ikebe, F V Brozovich
Apr 1, 1996·Journal of Muscle Research and Cell Motility·K MabuchiC L Wang
Feb 7, 1997·The Journal of Biological Chemistry·K HiranoD J Hartshorne
Feb 1, 1997·Journal of Muscle Research and Cell Motility·P J GallagherJ T Stull

❮ Previous
Next ❯

Related Concepts

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.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

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.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. 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.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.