共查询到20条相似文献,搜索用时 31 毫秒
1.
Antibodies directed toward PGF2α which cross react with PGE2 only slightly were used to detect conversion of PGE2 to PGF2α by homogenates of several rat tissues. This conversion by rat heart homogenates was demonstrated to be reversible, lost after trypsin digestion, and inhibited by several sulfhydryl blocking agents. The activity of the rat heart homogenate was precipitable by ammonium sulphate, was not dialyzable, and was 50% destroyed when the homogenate was incubated at 50° for 5 min. In the rat, the heart had the highest activity, followed by the kidney, brain, and liver. Negligible activity was found in smooth muscle, skeletal muscle, and whole blood of rat. 相似文献
2.
F. J. Otto Ursula Hacker J. Zante J. Schumann W. Göhde M. L. Meistrich 《Histochemistry and cell biology》1979,60(3):249-254
Summary Lactate dehydrogenase (LDH) isozyme composition and localization was determined in sections of skeletal, heart and smooth muscle by the mixed aggregation immunocytochemical method using first antibody directed against purified human LDH-A4 (M4) or LDH-B4 (H4) followed by the enzymes LDH-A4 and LDH-B4, respectively. An even distribution of the two monomers in all fibres was seen with heart muscle and smooth muscle. Heart muscle had a low concentration of A-monomers and a high concentration of B-monomers, whereas the smooth muscle had equal concentrations of the two monomers. In contrast, skeletal muscle from m. quadriceps femoris was found to be composed of two muscle fibre types, one containing mainly A-, the other mainly B-monomers. On the basis of succinate dehydrogenase activity it was shown that the red (type 1) fibres contain mainly B-monomers and the white (type 2) fibres mainly A-monomers of LDH. 相似文献
3.
E J Miller J E Finch E Chung W T Butler P B Robertson 《Archives of biochemistry and biophysics》1976,173(2):631-637
Solutions of native Type III collagen (chain composition, [α1(III)]3) exhibit a rapid and dramatic decrease in relative viscosity when incubated with trypsin. Cleavage products of the reaction were precipitated with ammonium sulfate and isolated in denatured form by molecular sieve chromatography. They were found to be comprised of: α1(III)-T1 (molecular weight, 71,000) derived from the NH2-terminal portion of the Type III molecule; and α1(III)-T2 (molecular weight, 24,000) from the COOH-terminal portion of the molecule. Determination of the amino acid sequence at the NH2-terminal portion of α1(III)-T2 as well as at the COOH-terminus of α(III)-T1 demonstrated that the products arose from specific cleavage of the type III molecule at an arginine-glycine bond corresponding to residues 780–781 in the repetitive triplet sequence of the α1(III) chain. The results suggest that the trypsin-susceptible bond in the native Type III collagen molecule resides in a region characterized by a relative lack of the normal collagen helicity. 相似文献
4.
《Process Biochemistry》2014,49(10):1767-1777
Type II pepsin soluble collagens (PSC) were isolated from skeletal and head bone of silvertip shark; and examined for their biochemical and structural properties. Among the raw materials, the protein content (8.99%) was high in skeletal bone and the ash content (28%) was high in head bone. After the collagen extraction, the raw materials contained higher amount of ash content ranging from 82 to 88%. The hydroxyproline content of skeletal and skeletal PSC (30 and 113 mg/g) was higher than those head and head PSC. Both collagens were composed of two different α-chains (α1- and α2-chains) and were characterized as type II collagen. Amino acid analysis of skeletal and head PSC indicated imino acid contents of 156 and 175 amino acid residues per 1000 residues, respectively. Similar, Fourier transform infrared spectra of SCII and HCII were observed, which suggested that the isolation process did not affect the secondary structure and molecular order of collagen, particularly the triple–helical structure. Denaturation temperature of skeletal PSC (31 °C) was higher than that of head PSC. SEM microstructure of the collagens depicted a porous, fibrillary and multi-layered structure. These results suggested that the PSC isolated from skeletal and head bone of silvertip shark were found to be suitable biomaterial in commercial applications as alternatives to mammalian collagen. 相似文献
5.
Comparative study of various methods for the extraction of free creatine and phosphocreatine from mouse skeletal muscle 总被引:1,自引:0,他引:1
H. H. Berlet 《Analytical biochemistry》1974,60(2):347-357
The efficacy of various media regarding the extraction of free creatine and phosphocreatine of mouse skeletal muscle was evaluated. In anesthetized animals tissue was quick-frozen in situ and removed by means of a modified Rongeur forceps cooled in liquid N2. Homogenization of muscle tissue in 1 m
EDTA in 50% (v/v) ethanol at −20°C, which was gradually diluted with ice-cold 0.4
perchloric acid to a final concentration of 0.3
perchloric acid in 12.5% ethanol proved to be the most suitable procedure regarding rapid handling of tissue samples, recovery of total creatine, and the ratio of phosphocreatine to total creatine. Phosphocreatine values as high as 78% of total creatine of skeletal muscle were thus obtained. Extraction of free creatine and phosphocreatine with concentrated ethanolic solutions (50–80%, v/v) was found to be incomplete apparently due to irreversible binding of creatine and phosphocreatine to protein precipitates. 相似文献
6.
《Bioscience, biotechnology, and biochemistry》2013,77(1):312-315
1α,25-dihydroxyvitamin D3 [1,25-(OH)2D3] phosphorylates the extracellular signal-regulated kinase (ERK), a member of the mitogen-activated protein kinase (MAPK) family, within 30 sec in primary cultured chick skeletal muscle cells. MAPK of HeLa cell lines, which had been stably transfected with a cDNA library derived from mRNA of chick skeletal muscle cells, was also rapidly phosphorylated by 1,25-(OH)2D3. These cell lines have the potential to be a good tool for further investigation of rapid non-genomic mechanism activated by 1,25-(OH)2D3. 相似文献
7.
Yunike Akasaka Masaki TsunodaTomomi Ogata Tomohiro IdeKoji Murakami 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2010,1801(10):1115-1122
Leptin administration has been shown to enhance muscle lipid oxidation in relation to the energy expenditure. Both long-form (Ob-RL) and short-form leptin receptors (Ob-RS) are expressed in skeletal muscle, but the role of Ob-RS is unclear. In the present study, the role of Ob-RS in leptin-induced lipid oxidation in skeletal muscles was investigated using primary murine myotubes from m/m and db/db mice. Primary myotubes were treated with leptin (0.1, 1, 10, 100 nM) for 24 h. Lipid oxidation was determined by 14CO2 production rate from [1-14C] palmitate. Leptin was found to increase lipid oxidation in a dose- and time-dependent manner in db/db myotubes as well as in m/m myotubes. Leptin significantly increased phosphorylation of JAK2 and STAT3 in both types of myotube. Leptin-induced lipid oxidation was abolished by STAT3 siRNA. To investigate the mechanism underlying leptin-induced lipid oxidation, the effects of pharmacological inhibitors were examined. JAK2 or p38 MAPK inhibitor suppressed leptin-induced lipid oxidation and decreased STAT3 phosphorylation in both types of myotube, respectively. Leptin significantly increased phosphorylation of p38 MAPK, and leptin-induced lipid oxidation was abolished by treatment with p38 MAPK siRNA in both types of myotube. These results suggest that leptin induces lipid oxidation in skeletal muscle through the JAK2/p38 MAPK/STAT3 signaling pathway via not only Ob-RL but also Ob-RS. 相似文献
8.
Steen Larsen Regitze Kraunsøe Martin Gram Erich Gnaiger Jørn W. Helge Flemming Dela 《Analytical biochemistry》2014
The number of studies on mitochondrial function is growing as a result of the recognition of the pivotal role of an intact mitochondrial function in numerous diseases. Measurements of oxygen consumption by the mitochondria in human skeletal muscle are used in many studies. There are several advantages of studying mitochondrial respiration in permeabilized fibers (Pfi), but the method requires a manual procedure of mechanical separation of the fiber bundles in the biopsy and chemical permeabilization of the cell membrane. This is time-consuming and subject to interpersonal variability. An alternative is to use a semiautomatic tool for preparation of a homogenate of the muscle biopsy. We investigated whether the PBI shredder is useful in preparing a muscle homogenate for measurements of mitochondrial respiratory capacity. The homogenate is compared with the Pfi preparation. Maximal respiratory capacity was significantly reduced in the homogenate compared with the Pfi from human skeletal muscle. A marked cytochrome c response was observed in the homogenate, which was not the case with the Pfi, indicating that the outer mitochondrial membrane was not intact. The mitochondria in the homogenate were more uncoupled compared with the Pfi. Manual permeabilization is an advantageous technique for preparing human skeletal muscle biopsies for respirometry. 相似文献
9.
Bettina A Gentry J. Andries Ferreira Amanda J. McCambridge Marybeth Brown Charlotte L. Phillips 《Matrix biology》2010,29(7):638-644
Exercise intolerance, muscle fatigue and weakness are often-reported, little-investigated concerns of patients with osteogenesis imperfecta (OI). OI is a heritable connective tissue disorder hallmarked by bone fragility resulting primarily from dominant mutations in the proα1(I) or proα2(I) collagen genes and the recently discovered recessive mutations in post-translational modifying proteins of type I collagen. In this study we examined the soleus (S), plantaris (P), gastrocnemius (G), tibialis anterior (TA) and quadriceps (Q) muscles of mice expressing mild (+/oim) and moderately severe (oim/oim) OI for evidence of inherent muscle pathology. In particular, muscle weight, fiber cross-sectional area (CSA), fiber type, fiber histomorphology, fibrillar collagen content, absolute, relative and specific peak tetanic force (Po, Po/mg and Po/CSA respectively) of individual muscles were evaluated. Oim/oim mouse muscles were generally smaller, contained less fibrillar collagen, had decreased Po and an inability to sustain Po for the 300-ms testing duration for specific muscles; +/oim mice had a similar but milder skeletal muscle phenotype. +/oim mice had mild weakness of specific muscles but were less affected than their oim/oim counterparts which demonstrated readily apparent skeletal muscle pathology. Therefore muscle weakness in oim mice reflects inherent skeletal muscle pathology. 相似文献
10.
J ChandraRajan 《Biochemical and biophysical research communications》1978,83(1):180-186
Types I and III collagens were solubilized from fetal human skin by limited digestion with pepsin and precipitated by dialysis against 0.02 M Na2HPO4. Heat denaturation of the collagens in 2 M guanidine-HCl, pH 7.5, resulted in the precipitation of the contaminant pepsin which could be removed by centrifugation. Renaturation of the denatured collagens by dialysis against deionized water at 22° for 2 hours selectively precipitated the type III collagen fibrils. Type I collagen remained in solution. The simplicity and high recovery (77%) make this a suitable approach for the rapid estimation of type III collagen in small tissue samples. 相似文献
11.
Val A. Fajardo Eric Bombardier Chris Vigna Tahira Devji Darin Bloemberg Daniel Gamu Anthony O. Gramolini Joe Quadrilatero A. Russell Tupling 《PloS one》2013,8(12)
Sarcolipin (SLN) and phospholamban (PLN) inhibit the activity of sarco(endo)plasmic reticulum Ca2+-ATPases (SERCAs) by reducing their apparent affinity for Ca2+. A ternary complex between SLN, PLN, and SERCAs results in super-inhibition of SERCA activity. Analysis of skeletal muscle homogenate has limited our current understanding of whether SLN and PLN regulate SERCA1a, SERCA2a, or both in skeletal muscle and whether SLN and PLN are co-expressed in skeletal muscle fibers. Biopsies from human vastus lateralis were analyzed through single fiber Western blotting and immunohisto/fluorescence staining to circumvent this limitation. With a newly generated SLN antibody, we report for the first time that SLN protein is present in human skeletal muscle. Addition of the SLN antibody (50 µg) to vastus lateralis homogenates increased the apparent Ca2+ affinity of SERCA (K
Ca, pCa units) (-Ab, 5.85 ± 0.02 vs. +Ab, 5.95 ± 0.02) and maximal SERCA activity (μmol/g protein/min) (-Ab, 122 ± 6.4 vs. +Ab, 159 ± 11) demonstrating a functional interaction between SLN and SERCAs in human vastus lateralis. Specifically, our results suggest that although SLN and PLN may preferentially regulate SERCA1a, and SERCA2a, respectively, physiologically they both may regulate either SERCA isoform. Furthermore, we show that SLN and PLN co-immunoprecipitate in human vastus lateralis homogenate and are simultaneously expressed in 81% of the fibers analyzed with Western blotting which implies that super-inhibition of SERCA may exist in human skeletal muscle. Finally, we demonstrate unequivocally that mouse soleus contains PLN protein suggesting that super-inhibition of SERCA may also be important physiologically in rodent skeletal muscle. 相似文献
12.
Summary The distribution of several extracellular matrix macromolecules was investigated at the myotendinous junction of adult chicken gastrocnemius muscle. Localization using monoclonal antibodies specific for 3 basal lamina components (type IV collagen, laminin, and a basement membrane form of heparan sulfate proteoglycan) showed strong fluorescent staining of the myotendinous junction for heparan sulfate proteoglycan and laminin, but not for type IV collagen. In addition, a strong fluorescent stain was observed at the myotendinous junction using a monoclonal antibody against the subunit of the chicken integrin complex (antibody JG 22). Neither fibronectin nor tenascin were concentrated at the myotendinous junction, but instead were present in a fibrillar staining pattern throughout the connective tissue which was closely associated with the myotendinous junction. Tenascin also gave bright fluorescent staining of tendon, but no detectable staining of the perimysium or endomysium. Type I collagen was observed throughout the tendon and in the perimysium, but only faintly in the endomysium. In contrast, type III collagen was present brightly in the endomysium and in the perimysium, but could not be detected in the tendon except when associated with blood vessels and in the epitendineum, which stained intensely. Type VI collagen was found throughout the tendon and in all connective tissue partitions of skeletal muscle. The results indicate that one or more molecules of the integrin family may play an important role in the attachment of muscle to the tendon. This interaction does not appear to involve extensive binding to fibronectin or tenascin, but may involve laminin and heparan sulfate proteoglycan. 相似文献
13.
C E Schwartz C G Hellerqvist L W Cunningham 《Biochemical and biophysical research communications》1979,90(1):240-246
The predominant collagenous protein secreted during the attachment of freshly trypsinized human foreskin fibroblasts was found to be Type I procollagen. Evidence is presented that both the α1 and α2 chains exhibit a 3-hydroxyproline/4-hydroxyproline ratio 4–5 fold higher than that of normal Type I collagen. These findings suggest that caution should be exercised in assigning an observed increase in the 3-hydroxyproline/4-hydroxyproline ratio to the synthesis of a basement membrane type collagen. 相似文献
14.
Mary E. Englert Mary Jane Landes Arnold L. Oronsky S.S. Kerwar 《Cellular immunology》1984,87(2):357-365
Intravenous administration of Type II collagen to rats prior to immunization with Type II collagen suppresses hind paw inflammation, humoral response to Type II collagen, and the severity of the arthritic lesion. Suppression of inflammation and its severity as well as the humoral response can also be induced by the prior intravenous administration of α1 (II) CB10 a cyanogen bromide peptide derived from Type II collagen. Suppression of arthritis is disease specific; intravenous administration of either Type II collagen or α1 (II) CB10 does not have an effect on adjuvant-induced arthritis. These studies indicate that structural determinants of α1 (II) CB10 (Mr, 30,000), a peptide located near the carboxy terminus of the collagen molecule, can induce suppression and suggest that these determinants may be responsible for the suppression of arthritis when Type II collagen is administered intravenously. 相似文献
15.
16.
Eun-Jeong Lee Josine M. De Winter Danielle Buck Jeffrey R. Jasper Fady I. Malik Siegfried Labeit Coen A. Ottenheijm Henk Granzier 《PloS one》2013,8(2)
The effect of the fast skeletal muscle troponin activator, CK-2066260, on calcium-induced force development was studied in skinned fast skeletal muscle fibers from wildtype (WT) and nebulin deficient (NEB KO) mice. Nebulin is a sarcomeric protein that when absent (NEB KO mouse) or present at low levels (nemaline myopathy (NM) patients with NEB mutations) causes muscle weakness. We studied the effect of fast skeletal troponin activation on WT muscle and tested whether it might be a therapeutic mechanism to increase muscle strength in nebulin deficient muscle. We measured tension–pCa relations with and without added CK-2066260. Maximal active tension in NEB KO tibialis cranialis fibers in the absence of CK-2066260 was ∼60% less than in WT fibers, consistent with earlier work. CK-2066260 shifted the tension-calcium relationship leftwards, with the largest relative increase (up to 8-fold) at low to intermediate calcium levels. This was a general effect that was present in both WT and NEB KO fiber bundles. At pCa levels above ∼6.0 (i.e., calcium concentrations <1 µM), CK-2066260 increased tension of NEB KO fibers to beyond that of WT fibers. Crossbridge cycling kinetics were studied by measuring ktr (rate constant of force redevelopment following a rapid shortening/restretch). CK-2066260 greatly increased ktr at submaximal activation levels in both WT and NEB KO fiber bundles. We also studied the sarcomere length (SL) dependence of the CK-2066260 effect (SL 2.1 µm and 2.6 µm) and found that in the NEB KO fibers, CK-2066260 had a larger effect on calcium sensitivity at the long SL. We conclude that fast skeletal muscle troponin activation increases force at submaximal activation in both wildtype and NEB KO fiber bundles and, importantly, that this troponin activation is a potential therapeutic mechanism for increasing force in NM and other skeletal muscle diseases with loss of muscle strength. 相似文献
17.
The effect of embryo extract on the types of collagen synthesized by cultured chick chondrocytes. 总被引:3,自引:0,他引:3
Growth of embryonic chick chondrocytes in dialyzed embryo extract results in both a change in morphology of the cells toward that of a fibroblast and a change in the type of collagen synthesized from the cartilage-specific Type II collagen (chain composition [α1(II)]3) to a mixture of Type I collagen (chain composition [α1(I)]2α2) and the Type I trimer (chain composition [α1(I)]3). Analyses after 6 days of growth in embryo extract show that the synthesis of only Type I collagen and the Type I trimer can be detected. However, on subculturing the cells to a low density and allowing a period of growth without embryo extract, colonies of chondrocytes reappear and the synthesis of Type II collagen apparently resumes. It is suggested that the observed changes represent a “modulation” in cell behavior, this being expressed not only by the morphological changes but also by changes in cell-specific protein synthesis as demonstrated by the changes in the type of collagen synthesized. 相似文献
18.
Simone M. Ward Margaret E. Fransen Dipak K. Dube Donald A. Fischman Larry F. Lemanski 《Cell and tissue research》1995,282(3):399-406
Of the several proteins located within sarcomeric A-bands, C-protein, a myosin binding protein (MyBP) is thought to regulate and stabilize thick filaments during assembly. This paper reports the characterization of C-protein isoforms in juvenile and adult axolotls, Ambystoma mexicanum, by means of immunofluorescent microscopy and Western blot analyses. C-protein and myosin are found specifically within the A-bands, whereas tropomyosin and -actin are detected in the I-bands of axolotl myofibrils. The MF1 antibody prepared against the fast skeletal muscle isoform of chicken C-protein specifically recognizes a cardiac isoform (Axcard1) in juvenile and adult axolotls but does not label axolotl skeletal muscle. The ALD66 antibody, which reacts with the C-protein slow isoform in chicken, localizes only in skeletal muscle of the axolotl. This slow axolotl isoform (Axslow) displays a heterogeneous distribution in fibers of dorsalis trunci skeletal muscle. The C315 antibody against the chicken C-protein cardiac isoform identifies a second axolotl cardiac isoform (Axcard2), which is present also in axolotl skeletal muscle. No C-protein was detected in smooth muscle of the juvenile and adult axolotl with these antibodies.This work was supported by NIH grants HL-32184 and HL-37702 and a grant-in-aid from the American Heart Association to L.F.L. 相似文献
19.
Isolation of types I and V collagens from carp muscle 总被引:3,自引:0,他引:3
K Sato R Yoshinaka M Sato Y Itoh Y Shimizu 《Comparative biochemistry and physiology. B, Comparative biochemistry》1988,90(1):155-158
1. The major constituent of carp intramuscular connective tissue was found to be Type I collagen. 2. A collagen homologous to Type V collagen of higher vertebrate was also isolated from carp muscle. 3. Relative portion of Type V collagen was higher in carp muscle than in mammalian muscles. 相似文献
20.
《Comparative biochemistry and physiology. B, Comparative biochemistry》1989,92(1):219-223
- 1.1. The collagen content in the abdominal muscle of seven species including shrimp, prawn, lobster and squilla varied among the species ranging from 1.1 to 6.2% of total tissue protein and the content in pereiopod and thoracic muscles of four species of crab varied ranging from 0.2 to 0.8%.
- 2.2. These results indicate that the musculature in flexible part comprises a high proportion of collagen.
- 3.3. The major collagen from the crustacean muscle was found to be similar to Type V collagen from the vertebrate muscle with respect to the solubility and amino acid composition.