Mammalian skeletal muscle C-protein: purification from bovine muscle, binding to titin and the characterization of a full-length human cDNA

1992 ◽  
Vol 102 (4) ◽  
pp. 769-778
Author(s):  
D.O. Furst ◽  
U. Vinkemeier ◽  
K. Weber
Keyword(s):  
Skeletal Muscle ◽  
Protein Sequencing ◽  
Protein Isoforms ◽  
Fast Method ◽  
Mammalian Skeletal Muscle ◽  
Terminal Sequence ◽  
C Protein ◽  
Human Sequence ◽  
Associated Proteins ◽  
Bovine Skeletal Muscle

We report a fast method for the isolation of homogeneous C-protein from bovine skeletal muscle. In electron micrographs C-protein appears as short rods with a relatively uniform length of about 50 nm. Protein sequencing shows a single N-terminal sequence. Radio-labelled C-protein strongly decorates titin II and myosin rods but not myosin heads. Binding to titin II is retained in preparations lacking titin-associated proteins. Antibodies to bovine C-protein were used to screen a lambda gt11 cDNA library constructed from fetal human skeletal muscle. Clone HC38 is 3833 bp long and encodes a protein of 1138 amino acid residues. The start of the predicted sequence fits the N-terminal sequence of the bovine protein. All partial sequences obtained from the bovine protein (348 residues) and the sequence deduced from a partial chicken cDNA (Einheber and Fischman, 1990) can be aligned along the human sequence. The sequences of human and chicken C-proteins share 50% identity and 70% similarity. Along the repeat patterns of the human protein the fibronectin (Fn)-like domains are better conserved than the immunoglobulin (Ig)-like domains. Regions of strong divergence between chicken fast C-protein and human slow C-protein may represent differences in C-protein isoforms.

The Adaptive Potential of Skeletal Muscle Fibers

2002 ◽  
Vol 27 (4) ◽  
pp. 423-448 ◽  
Author(s):  
Dirk Pette
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Muscle Fibers ◽  
Protein Isoforms ◽  
Mammalian Skeletal Muscle ◽  
Adaptive Potential ◽  
Hybrid Fibers ◽  
Neuromuscular Activity ◽  
Skeletal Muscle Fibers ◽  
Initial Location

Mammalian skeletal muscle fibers display a great adaptive potential. This potential results from the ability of muscle fibers to adjust their molecular, functional, and metabolic properties in response to altered functional demands, such as changes in neuromuscular activity or mechanical loading. Adaptive changes in the expression of myofibrillar and other protein isoforms result in fiber type transitions. These transitions occur in a sequential order and encompass a spectrum of pure and hybrid fibers. Depending on the quality, intensity, and duration of the alterations in functional demand, muscle fibers may undergo functional transitions in the direction of slow or fast, as well as metabolic transitions in the direction of aerobic-oxidative or glycotytic. The maximum range of possible transitions in either direction depends on the fiber phenotype and is determined by its initial location in the fiber spectrum. Key words: Ca-sequestering proteins, energy metabolism, fiber type transition, myofibrillar protein isofonns, myosin, neuromuscular activity

The isoforms of C protein and their distribution in mammalian skeletal muscle

1985 ◽  
Vol 6 (4) ◽  
pp. 487-505 ◽  
Author(s):  
G. K. Dhoot ◽  
M. C. Hales ◽  
B. M. Grail ◽  
S. V. Perry
Keyword(s):  
Skeletal Muscle ◽  
Mammalian Skeletal Muscle ◽  
C Protein

Immunohistochemical analysis of C-protein isoforms in cardiac and skeletal muscle of the axolotl, Ambystoma mexicanum

1995 ◽  
Vol 282 (3) ◽  
pp. 399-406 ◽  
Author(s):  
Simone M. Ward ◽  
Margaret E. Fransen ◽  
Dipak K. Dube ◽  
Donald A. Fischman ◽  
Larry F. Lemanski
Keyword(s):  
Skeletal Muscle ◽  
Immunohistochemical Analysis ◽  
Ambystoma Mexicanum ◽  
Protein Isoforms ◽  
C Protein

Coordinate changes in C protein and myosin expression during skeletal muscle hypertrophy

1994 ◽  
Vol 267 (2) ◽  
pp. C443-C449 ◽  
Author(s):  
K. M. McCormick ◽  
K. M. Baldwin ◽  
F. Schachat
Keyword(s):  
Skeletal Muscle ◽  
Muscle Hypertrophy ◽  
Peptide Mapping ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Thick Filament ◽  
Protein Isoforms ◽  
Plantaris Muscle ◽  
C Protein ◽  
Adult Rat

In this study, two new C protein isoforms in adult rat skeletal muscle were resolved using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These isoforms migrated between previously identified fast (Cf) and slow (Cs) C protein isoforms; hence they were named intermediate C proteins (Ci1 and Ci2). Cyanogen bromide peptide mapping and Western blotting showed that the intermediate isoforms were more similar to Cs than Cf. The distribution of specific C protein and myosin heavy chain (MHC) isoforms was highly correlated in several hindlimb muscles, suggesting that the expression of these two thick-filament proteins is coordinated. This notion was tested by determining whether specific C protein and MHC isoforms change in parallel during muscle hypertrophy. Eight weeks after ablation of its synergists, the overloaded plantaris muscle showed significant increases in type IIa MHC and intermediate C protein, with corresponding decreases in type IIb MHC and Cf protein. These results indicate that C protein expression is linked to MHC expression during plantaris muscle hypertrophy.

scholarly journals Localization of C-protein isoforms in chicken skeletal muscle: ultrastructural detection using monoclonal antibodies.

1984 ◽  
Vol 98 (4) ◽  
pp. 1514-1522 ◽  
Author(s):  
J E Dennis ◽  
T Shimizu ◽  
F C Reinach ◽  
D A Fischman
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibodies ◽  
Fiber Type ◽  
Ultrastructural Localization ◽  
Absolute Number ◽  
Protein Isoforms ◽  
C Protein ◽  
Thick Filaments ◽  
C Proteins ◽  
Chicken Skeletal Muscle

Monoclonal antibodies (McAbs) specific for the fast (MF-1) and slow (ALD-66) isoforms of C-protein from chicken skeletal muscle have been produced and characterized. Using these antibodies it was possible to demonstrate that skeletal muscles of varying fiber type express different isoforms of this protein and that in the posterior latissimus dorsi muscle both isoforms are co-expressed in the same myofiber (17, 18). Since we had shown that both isoforms were present in all sarcomeres, it was feasible to test whether the two isoforms co-distributed in the same 43-nm repeat within the A-band, thereby establishing a minimum number of C-proteins per repeat in the thick filaments. Here we describe the ultrastructural localization of C-protein in myofibers from three muscle types of the chicken using these same McAbs. We observed that although C-protein was present in a 43-nm repeat along the filaments in all three muscles, there were marked differences in the absolute number and position occupied by the different isoforms. Since McAbs MF-1 and ALD-66 decorated the same 43-nm repeats in the A-bands of the posterior latissimus dorsal muscle, we suggest that at least two C-proteins can co-localize at binding sites 43 nm apart along thick filaments of this muscle.

Immunohistochemical analysis of C-protein isoforms in cardiac and skeletal muscle of the axolotl, Ambystoma mexicanum

1995 ◽  
Vol 282 (3) ◽  
pp. 399-406
Author(s):  
Simone M. Ward ◽  
Margaret E. Fransen ◽  
Dipak K. Dube ◽  
Donald A. Fischman ◽  
Larry F. Lemanski
Keyword(s):  
Skeletal Muscle ◽  
Immunohistochemical Analysis ◽  
Ambystoma Mexicanum ◽  
Protein Isoforms ◽  
C Protein

scholarly journals The major myosin-binding domain of skeletal muscle MyBP-C (C protein) resides in the COOH-terminal, immunoglobulin C2 motif.

1993 ◽  
Vol 123 (3) ◽  
pp. 619-626 ◽  
Author(s):  
T Okagaki ◽  
F E Weber ◽  
D A Fischman ◽  
K T Vaughan ◽  
T Mikawa ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Amino Acids ◽  
Variable Number ◽  
Terminal Sequence ◽  
C Protein ◽  
C2 Domains ◽  
Type Iii ◽  
Myosin Filaments ◽  
Fibronectin Type Iii ◽  
Myosin Binding

A common feature shared by myosin-binding proteins from a wide variety of species is the presence of a variable number of related internal motifs homologous to either the Ig C2 or the fibronectin (Fn) type III repeats. Despite interest in the potential function of these motifs, no group has clearly demonstrated a function for these sequences in muscle, either intra- or extracellularly. We have completed the nucleotide sequence of the fast type isoform of MyBP-C (C protein) from chicken skeletal muscle. The deduced amino acid sequence reveals seven Ig C2 sets and three Fn type III motifs in MyBP-C. alpha-chymotryptic digestion of purified MyBP-C gives rise to four peptides. NH2-terminal sequencing of these peptides allowed us to map the position of each along the primary structure of the protein. The 28-kD peptide contains the NH2-terminal sequence of MyBP-C, including the first C2 repeat. It is followed by two internal peptides, one of 5 kD containing exclusively spacer sequences between the first and second C2 motifs, and a 95-kD fragment containing five C2 domains and three fibronectin type III motifs. The C-terminal sequence of MyBP-C is present in a 14-kD peptide which contains only the last C2 repeat. We examined the binding properties of these fragments to reconstituted (synthetic) myosin filaments. Only the COOH-terminal 14-kD peptide is capable of binding myosin with high affinity. The NH2-terminal 28-kD fragment has no myosin-binding, while the long internal 100-kD peptide shows very weak binding to myosin. We have expressed and purified the 14-kD peptide in Escherichia coli. The recombinant protein exhibits saturable binding to myosin with an affinity comparable to that of the 14-kD fragment obtained by proteolytic digestion (1/2 max binding at approximately 0.5 microM). These results indicate that the binding to myosin filaments is mainly restricted to the last 102 amino acids of MyBP-C. The remainder of the molecule (1,032 amino acids) could interact with titin, MyBP-H (H protein) or thin filament components. A comparison of the highly conserved Ig C2 domains present at the COOH-terminus of five MyBPs thus far sequenced (human slow and fast MyBP-C, human and chicken MyBP-H, and chicken MyBP-C) was used to identify residues unique to these myosin-binding Ig C2 repeats.

Size and charge heterogeneity of C-protein isoforms in avian skeletal muscle. Expression of six different isoforms in chicken muscle

1989 ◽  
Vol 10 (5) ◽  
pp. 369-378 ◽  
Author(s):  
H. Takano-Ohmuro ◽  
S. M. Goldfine ◽  
T. Kojima ◽  
T. Obinata ◽  
D. A. Fischman
Keyword(s):  
Skeletal Muscle ◽  
Protein Isoforms ◽  
Charge Heterogeneity ◽  
C Protein ◽  
Chicken Muscle

scholarly journals Calcium Ion in Skeletal Muscle: Its Crucial Role for Muscle Function, Plasticity, and Disease

2000 ◽  
Vol 80 (3) ◽  
pp. 1215-1265 ◽  
Author(s):  
Martin W. Berchtold ◽  
Heinrich Brinkmeier ◽  
Markus Müntener
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Binding Proteins ◽  
Muscle Fibers ◽  
Protein Isoforms ◽  
Muscle Performance ◽  
High Plasticity ◽  
Mammalian Skeletal Muscle ◽  
Variable Expression ◽  
Functional Features

Mammalian skeletal muscle shows an enormous variability in its functional features such as rate of force production, resistance to fatigue, and energy metabolism, with a wide spectrum from slow aerobic to fast anaerobic physiology. In addition, skeletal muscle exhibits high plasticity that is based on the potential of the muscle fibers to undergo changes of their cytoarchitecture and composition of specific muscle protein isoforms. Adaptive changes of the muscle fibers occur in response to a variety of stimuli such as, e.g., growth and differentition factors, hormones, nerve signals, or exercise. Additionally, the muscle fibers are arranged in compartments that often function as largely independent muscular subunits. All muscle fibers use Ca2+ as their main regulatory and signaling molecule. Therefore, contractile properties of muscle fibers are dependent on the variable expression of proteins involved in Ca2+ signaling and handling. Molecular diversity of the main proteins in the Ca2+ signaling apparatus (the calcium cycle) largely determines the contraction and relaxation properties of a muscle fiber. The Ca2+ signaling apparatus includes 1) the ryanodine receptor that is the sarcoplasmic reticulum Ca2+ release channel, 2) the troponin protein complex that mediates the Ca2+ effect to the myofibrillar structures leading to contraction, 3) the Ca2+pump responsible for Ca2+ reuptake into the sarcoplasmic reticulum, and 4) calsequestrin, the Ca2+storage protein in the sarcoplasmic reticulum. In addition, a multitude of Ca2+-binding proteins is present in muscle tissue including parvalbumin, calmodulin, S100 proteins, annexins, sorcin, myosin light chains, β-actinin, calcineurin, and calpain. These Ca2+-binding proteins may either exert an important role in Ca2+-triggered muscle contraction under certain conditions or modulate other muscle activities such as protein metabolism, differentiation, and growth. Recently, several Ca2+signaling and handling molecules have been shown to be altered in muscle diseases. Functional alterations of Ca2+ handling seem to be responsible for the pathophysiological conditions seen in dystrophinopathies, Brody's disease, and malignant hyperthermia. These also underline the importance of the affected molecules for correct muscle performance.

Distribution of c-protein isoforms in sarcomeres of developing chick skeletal muscle

1988 ◽  
Vol 46 ◽  
pp. 226-227
Author(s):  
D. A. Fischman ◽  
J. E. Dennis ◽  
T. Obinata ◽  
H. Takano-Ohmuro
Keyword(s):  
Skeletal Muscles ◽  
Thick Filament ◽  
Protein Isoforms ◽  
Actin Binding ◽  
Striated Muscles ◽  
C Protein ◽  
Sarcomeric Protein ◽  
Thick Filaments ◽  
Cross Bridges ◽  
Bridge Bearing

C-protein is a 150 kDa protein found within the A bands of all vertebrate cross-striated muscles. By immunoelectron microscopy, it has been demonstrated that C-protein is distributed along a series of 7-9 transverse stripes in the medial, cross-bridge bearing zone of each A band. This zone is now termed the C-zone of the sarcomere. Interest in this protein has been sparked by its striking distribution in the sarcomere: the transverse repeat between C-protein stripes is 43 nm, almost exactly 3 times the 14.3 nm axial repeat of myosin cross-bridges along the thick filaments. The precise packing of C-protein in the thick filament is still unknown. It is the only sarcomeric protein which binds to both myosin and actin, and the actin-binding is Ca-sensitive. In cardiac and slow, but not fast, skeletal muscles C-protein is phosphorylated. Amino acid composition suggests a protein of little or no αhelical content. Variant forms (isoforms) of C-protein have been identified in cardiac, slow and embryonic muscles.

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