Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Treatment of relaxed skinned rabbit psoas muscle fibers with 0.1 mM N-phenylmaleimide (NPM) for 1 h locks all of the crossbridges in a weakly-binding state resembling that of the myosin.ATP crossbridge. Under these conditions, NPM reacts mainly with myosin heavy chain (Barnett et al. (1992) Biophys. J. 61, 358-367). Here the specific sites for that reaction are explored. Small bundles of rabbit psoas muscle fibers were treated with Triton X-100 to make the fiber sarcolemmas permeable. The bundles were treated with 0.1 mM [14C]NPM for 1 h, and homogenized for SDS-PAGE. 43 +/- 2.2% of the muscle fiber protein ran in the myosin heavy chain band, the same as for untreated fibers. An alkylating stoichiometry of 2.2 +/- 0.33 moles NPM per mole myosin heavy chain was determined. Exhaustive trypsin digestion followed by two-dimensional thin-layer chromatography and reverse-phase HPLC revealed two major sites on myosin heavy chain for NPM binding. The sites contained about the same amount of linked NPM, suggesting that the reaction stoichiometry of each site under the conditions studied is approx. 1 mol NPM/mol myosin heavy chain. Comparison of the labeled tryptic peptides with NPM-reacted synthetic SH1 and SH2 tryptic peptides and analysis of the treated fiber bundles' ATPase activity suggested that the sites for NPM reaction on myosin heavy chain when it locks crossbridges in a weakly-binding state are Cys-697 (SH2) and Cys-707 (SH1).
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PMID:The site and stoichiometry of the N-phenylmaleimide reaction with myosin when weakly-binding crossbridges are formed in skinned rabbit psoas fibers. 749 34

The present study investigates the effects of chronic low-frequency electrical stimulation on the external urethral sphincter (EUS) of rabbits through the biochemical analysis of the isoforms of myosin light and heavy chains. Twenty-eight adult male rabbits were used in the test. Of these, the EUS of 14 rabbits were continuously stimulated directly at 1.8 Hz, 0.8 msec, 5.0 V with an implantable electrical cardiac pacemaker for more than 10 weeks. The other 14 rabbits were used as the control group and were nurtured under the same conditions as the stimulated group but without the electrical stimulation. Upon conclusion of the stimulation program, the urethra was removed from all 28 rabbits. The portion of the urethra containing the EUS (from the middle of the prostate to the pelvic diaphragm) was cut transversely into thin serial sections and glycerinated. The glycerinated muscle fibers were then isolated under a stereomicroscope and samples for electrophoretic analysis were prepared. Two-dimensional electrophoresis according to the procedure of O'Farrell for myosin light chains and SDS-PAGE containing 40% glycerol for myosin heavy chains were carried out. The molar ratio of myosin subunits was determined by quantification through the dye elution process. The average percentages of slow myosin light chain molecules in 8 unstimulated and 8 stimulated EUS were 33.4 +/- 8.9% (mean +/- SD) and 70.1 +/- 12.8%, respectively. The average percentages of slow myosin heavy chain molecules in 6 unstimulated and 6 stimulated EUS were 17.6 +/- 5.7% and 40.2 +/- 7.1%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Effects of chronic low-frequency electrical stimulation on the external urethral sphincter of male rabbits--electrophoretic analyses of myosin light and heavy chain isoforms]. 763 39

Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE), glycerol SDS-PAGE, two-dimensional electrophoresis, and protein immunoblotting techniques were used to identify myosin heavy chain (MHC) and light chain (MLC) isoforms in limb and masticatory muscles of the cat and American opossum. The fibre types in which these isoforms are expressed were identified by histochemistry and immunohistochemistry. Antibodies specific for the type IIM MHC isoform characteristic of cat jaw-closing muscles and the type I MHC isoform were produced and characterized. The IIM antibody stained the majority of fibres found in the jaw-closing muscles of both species. These IIM-containing fibres characteristically had a histochemical ATPase that remained active after both acid and alkali pre-incubations. A minority of type I fibres was also present in cat jaw-closing muscles, and these reacted positively with antibody specific for type I MHC. It was confirmed that the vast majority of fibres in the cat jaw-closing muscles contained only the characteristic masticatory MHC (IIM) and masticatory MLCs (LC1m and LC2m). These muscles did not contain either the type II fibre isoforms of limb muscles or the atrial cardiac (alpha-cardiac) MHC. The type IIM MHC could also be identified in jaw-closing muscles of the opossum. Two-dimensional gel electrophoresis was used to identify the MLC composition of single, histochemically defined, type I fibres in the cat soleus and deep masseter. The type I fibres of limb muscle contained the usual slow MLCs, but type I fibres from the jaw-closing muscles contained only the masticatory light chains.
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PMID:Myosin expression in the jaw-closing muscles of the domestic cat and American opossum. 763 44

During growth and development, dietary intake changes from being predominantly liquid in the newborn period to mixed solid liquid meals. These alterations in diet vary the functional demands placed on the stomach. It has been shown that, during development, smooth muscle of the stomach undergoes changes in the mechanism responsible for the contractile process. In this study, we have investigated the possibility that there are structural changes in two of the major proteins that are responsible for generation of force during smooth muscle contraction: actin and myosin. Actin and myosin were identified in newborn kittens (1 wk old) and adult gastric smooth muscle using one-dimensional SDS-PAGE. Although both the antrum and fundus of the kitten have significantly smaller total amounts of actin and myosin per mg protein than the adult, the ratio of actin to myosin is not significantly different between the age groups. Two different myosin heavy chain (MHC) isoforms, MHC1 (205 kD) and MHC2 (200 kD), were identified in all tissues. The relative amount of MHC1 remained constant during maturation of the stomach. We observed an increase in the amount of MHC2 in the adult, which resulted in a decreased ratio of MHC1 to MHC2 in the adult. We postulate that the decreased quantity of actin and myosin in the kitten stomach and the observed changes in the ratio of the MHC isoforms are related to changes in the gastric motor that occur during growth and development.
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PMID:Characterization of actin and myosin in the developing stomach. 773 58

A tissue-type transglutaminase (TGase) was purified from liver tissue of the red sea bream, Pagrus major, by ion-exchange chromatography and heparin-Sepharose affinity chromatography. Its activity was assessed using a fluorometric assay to measure the incorporation of monodansylcadaverine into N,N'-dimethyl casein. The molecular mass of purified TGase was estimated to be 78 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme required Ca2+ to express its activity, although 10 mM Sr2+ also activated the enzyme fully. TGase activity was maximal at pH 9.0-9.5, and the enzyme was strongly inhibited by sulfhydryl reagents. The purified enzyme catalyzed the cross-linking of myosin heavy chain obtained from Alaska pollack, resulting in gelation of an actomyosin solution. The partial amino acid sequence of this fish TGase showed divisionally significant similarity to TGase from guinea pig liver.
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PMID:Purification and characterization of a tissue-type transglutaminase from red sea bream (Pagrus major). 776 97

Myosin heavy chain composition of a large number (288) of single fibres from slow (soleus), and fast (superficial part of tibialis anterior, and plantaris) muscles of adult (3-5-month-old) Wistar rats was determined. A combination of SDS-PAGE and monoclonal antibodies against myosin heavy chains allowed to identify four myosin heavy chain isoforms (1, 2A, 2X, and 2B) and to detect myosin heavy chain coexistence. Four groups of fibres containing only one myosin heavy chain (1 myosin heavy chain, 2A myosin heavy chain, 2X myosin heavy chain, and 2B myosin heavy chain), and five groups containing more than one myosin heavy chain (1 and 2A myosin heavy chains, 2A and 2X myosin heavy chains, 2X and minor amounts of 2B (2X-2B fibres), 2B and minor amounts of 2X (2B-2X fibres), and 2A, 2X, and 2B myosin heavy chain were identified and their relative percentages were assessed. Coexistence of fast myosin heavy chain isoforms was found to be very frequent (50% of the fibres in plantaris, and 30% in tibialis anterior), whereas coexistence of slow and fast (2A) myosin heavy chain was very rare. Maximum shortening velocity (V0) was determined using the slack-test procedure in a subset of 109 fast fibres from the above population. The values of V0 formed a continuum extending from 2A to 2X to 2X-2B to 2B-2X to 2B fibres. 2A fibres had the lowest value of V0 and 2B fibres the highest. Only the differences between 2A and 2B and 2A and 2B-2X fibres were statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Maximum shortening velocity and coexistence of myosin heavy chain isoforms in single skinned fast fibres of rat skeletal muscle. 780 35

The effects of 1, 2, and 3 wk of hindlimb suspension (HS) on force-velocity and power characteristics of single rat soleus fibers were determined. After 1, 2, or 3 wk of HS, small fiber bundles were isolated, placed in skinning solution, and stored at -20 degrees C until studied. Single fibers were isolated and placed between a motor arm and force transducer, functional properties were studied, and fiber protein content was subsequently analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Additional fibers were isolated from soleus of control and after 1 and 3 wk of HS, and fiber type distribution and myosin light chain stoichiometry were determined from SDS-PAGE analysis. After 1 wk of HS, percent type I fibers declined from 82 to 74%, whereas hybrid fibers increased from 10 to 18%. Percent fast type II fibers increased from 8% in control and 1 wk of HS to 26% by 3 wk of HS. Most fibers showed an increased unloaded maximal shortening velocity (Vo), but myosin heavy chain remained entirely slow type I. The mechanism for increased Vo is unknown. There was a progressive decrease in fiber diameter (14, 30, and 38%) and peak force (38, 56, and 63%) after 1, 2, and 3 wk of HS, respectively. One week of HS resulted in a shift of the force-velocity curve, and between 2 and 3 wk of HS the curve shifted further such that Vo was higher than control at all relative loads < 45% peak isometric force. Peak absolute power output of soleus fibers progressively decreased through 2 wk of HS but showed no further change at 3 wk.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Force-velocity and power characteristics of rat soleus muscle fibers after hindlimb suspension. 783 76

Human masseter muscle is highly unusual since it contains relatively large numbers of fibres with variable myofibrillar ATPase staining as well as fibres that express neonatal and alpha-cardiac myosin heavy chain (MHC). These findings however, have not been organised together into a fibre type classification scheme. Biopsies from the anterior superficial area of masseter were collected from a large sample of healthy young adults. Biopsies were sectioned and stained for myofibrillar ATPase reactivity and the presence of MHC isoforms as detected by a series of antibodies. The MHC composition of the same biopsies was also analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). A series of rectus abdominis muscle biopsies were analysed similarly to serve as a control for type I, IIA and IIB fibres and isoforms. From the histochemical, immunohistochemical and biochemical experiments we found the masseter to contain type I, IM, IIC, IIA and IIB fibres as previously classified, but in addition there were type neonatal, alpha-cardiac, and 'other' (three or more myosins including neonatal and alpha-cardiac). The percentage of each fibre type was highly variable in masseter biopsies, but generally type I fibres were most common, and the proportion of IIB, neonatal, alpha-cardiac and 'other' fibres was low. Even in biopsies that contained relatively large amounts of these last three fibre types, the amount of neonatal and/or alpha-cardiac MHC detected on SDS-PAGE was limited, suggesting that these MHCs are a minor component in the fibres in which they are expressed.
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PMID:Fibre type classification and myosin isoforms in the human masseter muscle. 783 42

The goal of the current investigation was to characterize, purify, and identify the proteins that bind surfactant protein A (SP-A). Several polypeptides were purified by SP-A affinity chromatography, and the 200 kD major polypeptide that reacted with SP-A on ligand blots was purified further by preparative SDS-PAGE. Protein sequencing of proteolytically derived subfragments of this polypeptide gave sequences that corresponded completely with nonmuscle (cellular) myosin heavy chain. The 200 kD polypeptide was then found to be immunoreactive with antibodies against cellular myosin. A smaller polypeptide of 135 kD also binds SP-A and appears to be a proteolytic fragment of the 200 kD peptide. The ability of SP-A to bind myosin was confirmed in a microtiter well assay and was found to be concentration dependent. We speculated that the physiologic relevance of the interaction of SP-A with myosin might be to facilitate clearance of myosin from the alveolar subphase following its release during lung injury. In support of this hypothesis, we found that there were detectable levels of myosin in lavage fluid and that SP-A could indeed enhance uptake and degradation of myosin by alveolar macrophages.
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PMID:Interaction of surfactant protein A with cellular myosin. 794 98

We studied the difference in the proportion of fast and slow myosin subunits in axially subdivided external urethral sphincter of male rabbit using myosin light chain and heavy chain analyses. The whole urethras from 6 adult male Japanese White rabbits were sagittally bisected and one halves from all animals were processed for myosin light chain analysis and another halves from all animals for myosin heavy chain analysis. Each half urethra containing the external urethral sphincter was subdivided into 4 parts, namely prostatic (P), prostatic apical (PA), infraprostatic (IP) and bullbourethral glandular (BUG) portions, from cranial to caudal direction. The electrophoretic samples from 4 different parts were separately processed. The external urethral sphincter muscle in each part from all animals were processed together for a sample because of the minute amount of the muscle. Two-dimensional electrophoresis according to the procedure of O'Farrell for myosin light chains and SDS-polyacrylamide gel electrophoresis containing 40% glycerol for myosin heavy chains were carried out. Molar ratio of myosin subunits were estimated by the dye elution procedure. The relative proportions of slow myosin molecules in respective parts were 33.4% (P), 26.3% (PA), 18.5% (IP) and 11.0% (BUG) from myosin light chain analysis and 20.3% (P), 16.1% (PA), 7.2% (IP) and 5.0% (BUG) from myosin heavy chain analysis. The urethral striated musculature in male rabbit was predominantly composed of fast myosin subunits as a whole. But the proportion of slow myosin subunits occupied a relatively high percentage in the proximal region and tended to decrease toward the distal end.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Biochemical analysis of the external striated urethral sphincter of male rabbits. Difference in the proportions of muscle fiber types in the male rabbit external urethral sphincter by axial subdivisional study]. 799 Mar 2


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