Pulse Electrophoresis of Muscle Myosin Heavy Chains in Sodium Dodecyl Sulfate–Polyacrylamide Gels

2001 ◽  
Vol 291 (2) ◽  
pp. 229-236 ◽  
Author(s):  
José A.A. Sant'Ana Pereira ◽  
Marion Greaser ◽  
Richard L. Moss
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Myosin Heavy Chains ◽  
Polyacrylamide Gels ◽  
Heavy Chains ◽  
Dodecyl Sulfate ◽  
Muscle Myosin

Two different heavy chains are found in smooth muscle myosin

1986 ◽  
Vol 250 (6) ◽  
pp. C861-C870 ◽  
Author(s):  
A. S. Rovner ◽  
M. M. Thompson ◽  
R. A. Murphy
Keyword(s):  
Smooth Muscle ◽  
Sodium Dodecyl Sulfate ◽  
Heavy Chain ◽  
Sodium Dodecyl ◽  
Smooth Muscles ◽  
Protein Band ◽  
Smooth Muscle Myosin ◽  
Heavy Chains ◽  
Dodecyl Sulfate ◽  
Muscle Myosin

Two putative myosin heavy chains designated SM1 and SM2 were detected on a 3.5% polyacrylamide-sodium dodecyl sulfate gel electrophoresis system loaded with homogenates of several mammalian smooth muscles. The two polypeptides were present in nearly equal amounts in all smooth muscle tissues tested and in myosin purified from swine carotid media and stomach. Both proteins were equally stained by smooth muscle-specific myosin antibodies. The smaller of the polypeptides had a mobility nearly identical to that of the single heavy chain observed in purified fast-twitch skeletal myosin. Electrophoresis of pyrophosphate extracts from swine carotid media, swine stomach, rabbit thoracic aorta, and guinea pig taenia coli on nondenaturing pyrophosphate gels revealed a single protein band. When subsequently electrophoresed on a sodium dodecyl sulfate gel, the native bands from swine tissue extracts revealed the two putative heavy chains in nearly equal amounts, as well as a large amount of a higher molecular weight peptide whose properties reflect those of filamen. Sodium dodecyl sulfate gel analysis of the myosin band from pyrophosphate gels of purified swine stomach myosin showed exclusively the two heavy chains in a nearly 1:1 ratio. Smooth muscle myosin migrates homogeneously on pyrophosphate gels, and the virtual equality of the two heavy chains may reflect the presence of large amounts of a myosin isoenzyme, which is a heavy-chain heterodimer.

Smooth muscle myosin heavy chains combine to form three native myosin isoforms

1993 ◽  
Vol 264 (5) ◽  
pp. H1653-H1662 ◽  
Author(s):  
A. E. Tsao ◽  
T. J. Eddinger
Keyword(s):  
Smooth Muscle ◽  
Sodium Dodecyl ◽  
Coiled Coil ◽  
Smooth Muscle Myosin ◽  
Polyacrylamide Gel ◽  
Myosin Isoforms ◽  
Myosin Heavy Chains ◽  
Polyacrylamide Gels ◽  
Heavy Chains ◽  
Muscle Myosin

Two smooth muscle myosin heavy chains (MHC; SM1 and SM2) of approximately 204 and 200 kDa exist in smooth muscle cells and can be visualized on reducing sodium dodecyl sulfate (SDS)-polyacrylamide gels. Chymotryptic digestion of the native myosin molecule results in two fragments: heavy meromyosin (HMM) and light meromyosin (LMM). LMM is the alpha-helical coiled-coil carboxy terminal half of the molecule containing the difference peptide between SM1 and SM2. Electrophoresis of the LMM fragments on a reducing SDS-polyacrylamide gel resolves two subunits from the two MHC [LM1 from SM1 (approximately 100 kDa) and LM2 from SM2 (approximately 95 kDa), where LM1 and LM2 are LMM from SM1 and SM2, respectively]. CuCl2 oxidation of the LMM fragment forms intramolecular disulfide bonds between adjacent cysteines on the two LMM fragments. When the native LMM is oxidized with CuCl2 and run on a nonreducing SDS-polyacrylamide gel, three bands are observed, which migrate at approximately 195, 190, and 185 kDa (bands 1, 2, and 3). Excision of these bands and electrophoresis on a reducing SDS-polyacrylamide gel show their subunit composition. Band 1 is composed solely of LM1. Band 2 is composed of an equal ratio of LM1 and LM2, and band 3 is composed solely of LM2. Using a variety of biochemical procedures, along with nonreducing SDS-polyacrylamide gels, we interpret these results to indicate that there are three smooth muscle myosin isoforms that result from the various combinations of the two smooth muscle MHC (SM1 homodimer, SM1-SM2 heterodimer, and SM2 homodimer).

Efficacy and compatibility with mass spectrometry of methods for elution of proteins from sodium dodecyl sulfate–polyacrylamide gels and polyvinyldifluoride membranes

2004 ◽  
Vol 330 (1) ◽  
pp. 87-97 ◽  
Author(s):  
Charlotte Sværke Jørgensen ◽  
Mitchell Jagd ◽  
Birgitte Kjær Sørensen ◽  
Jim McGuire ◽  
Vibeke Barkholt ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gels ◽  
Dodecyl Sulfate

Detection of N-glycans on small amounts of glycoproteins in tissue samples and sodium dodecyl sulfate–polyacrylamide gels

2012 ◽  
Vol 423 (2) ◽  
pp. 253-260 ◽  
Author(s):  
Takeshi Yoshimura ◽  
Gen Yamada ◽  
Mai Narumi ◽  
Takako Koike ◽  
Akihiro Ishii ◽  
...  
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gels ◽  
Tissue Samples ◽  
Dodecyl Sulfate
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