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R Starr

Publications and source records attributed to R Starr.

At least 55 records · Page 3Linked to original sources
PubMed1988

Phosphofructokinase: a component of the thick filament?

F-protein, a consistent contaminant of myosin preparations, has been shown to be phosphofructokinase, the key regulatory enzyme of glycolysis. In homogenates of rigor muscle most of the phosphofructokinase sediments with the myofibrils, suggesting that in the living muscle cell phosphofructokinase is not in the soluble fraction as was formerly thought, but bound to the myofibrils. Fluorescent antibody to F-protein labels myofibrils in a zone in each half of the A-band. The increase in separation of the zones across the A-band with increase in sarcomere length suggests that the antibody binds to the parts of the cross-bridge regions of the thick filament within the H-zone. It therefore seems likely that phosphofructokinase is located in the cross-bridge region of the thick filament, but that access of antibody is restricted by overlapping thin filaments.

Animals↗
PubMed1986

The ultrastructural location of C-protein, X-protein and H-protein in rabbit muscle.

Purified antibodies to the thick filament accessory proteins, C-protein, X-protein and H-protein, have been used to label fibres of three rabbit muscles, psoas (containing mainly fast white fibres), soleus (containing mainly slow red fibres) and plantaris (a muscle of mixed fibre type) and their location has been examined by electron microscopy. These accessory proteins are present on one or more of a set of eleven transverse stripes about 43 nm apart that have been observed previously in each half A-band. Each protein has a limited set of characteristic distributions. H-protein is present on stripe 3 (counting from the M-line) in the majority of psoas fibres but is absent in soleus and plantaris muscle. C-protein can occur on stripes 4-11 (the commonest pattern seen in psoas); on stripes 5-11 (in psoas and plantaris); on stripes 3 together with stripes 5-11 (in plantaris); or on none (in red fibres of all three muscles). X-protein can occur on stripes 3-11 in the red fibres of all three muscles; on stripe 4 only (in psoas and plantaris); on stripes 3 and 4 (in psoas and plantaris) or on none. Stripes labelled with anti-X are wider than those labelled with anti-C and consist of a doublet with an internal spacing of 16 nm. The patterns for the three accessory proteins, while overlapping, are in no case identical; this suggests the proteins do not simply substitute for one another. The precise axial positions of the anti-C labelled stripes differ from those of the anti-X stripes; the anti-X stripes lie about 8-9 nm further from the M-line than the corresponding anti-C stripes. This implies that the inner member of an X-protein doublet lies in a very similar position to a C-protein stripe. The anti-H labelled stripe seen in most psoas fibres lies 14 nm nearer the M-line than stripe 3 of the anti-X labelled array in psoas red fibres and is staggered from a continuation of the C-protein array by about 4 nm. The labelling patterns were constant within a fibre and suggest a very precise assembly mechanism. The number of classes of fibre, as defined by the accessory proteins present and their arrangement, exceeds the number of fibre types presently recognized.

Animals↗
PubMed1985

The structure of C-protein and X-protein molecules and a polymer of X-protein.

C-protein and X-protein are components of the thick filaments in vertebrate skeletal muscles and occupy similar locations in different fibre types. We find that the molecules are both rods about 30 to 40 A wide, but they differ significantly in their lengths, the X-protein molecule being about 350 A long and the C-protein molecule about 280 A. This suggests they are not isoforms. The short length of the C-protein molecule implies that it cannot act in the thick filament as a length-determining agent by a simple vernier mechanism. X-protein associates at low ionic strength (KCl concentration less than 0.07 M) but, unlike C-protein, forms long ordered polymers. These have been examined by electron microscopy to gain information on the molecular shape and on how the molecules interact. The polymers are helically twisted ribbons with a repeat distance along the axis of 660 A. The cross-section of the ribbon is approximately elliptical with major and minor axes of 405 A and 166 A, respectively. From an analysis of the micrographs by optical diffraction, we deduce that the molecules run across the face of the ribbon at an angle of about 15 degrees to the diameter and lie on a two-stranded helix. Models for the polymer are discussed in which the molecules are slightly bowed outwards and bind to each other only at their ends. We suggest that interactions similar to those in the polymer might occur in the thick filaments of muscle, and propose that at each axial position where X-protein attaches along the myosin filament, three X-protein molecules might form an approximately triangular ring around the filament backbone. The appearance of the X-protein polymers is similar to that of the twisted structures called paired helical filaments that make up the neurofibrillary tangles associated with dementia of the Alzheimer type.

Animals↗
PubMed1985

Location of C-protein, H-protein and X-protein in rabbit skeletal muscle fibre types.

The locations of C-protein, H-protein and X-protein in rabbit psoas, plantaris and soleus muscles have been investigated with fluorescently tagged specific antibodies. Two systems have been examined: isolated myofibrils allowed the locations of these proteins within the sarcomere to be determined, while cryosections allowed a comparison of the amounts of these proteins between different types of fibre in the three muscles. Using antibody-labelled cryosections, we find that the amounts of each of these proteins depends closely on the fibre type. In all the muscles studied, C-protein is present in the largest amounts in fast white and fast intermediate fibres and is absent from slow red fibres, while X-protein is absent from fast white fibres and is present in the largest amounts in fast and slow red fibres. In psoas muscle, H-protein is present in the largest amounts in fast white fibres and is absent in fast and slow red fibres. In plantaris muscle, however, H-protein is absent from fast white fibres but occurs in some slow red fibres. All psoas myofibrils label with anti-C and anti-H and a minority label with anti-X. In each case the pattern of labelling is a zone in each half of the A-band. Measured across the middle of the A-band, the zones for H-protein are much closer together than for C-protein; the centre-to-centre spacings are 0.35 micron for anti-H and 0.64 micron for anti-C. The fluorescent zones for X-protein are slightly but significantly closer (0.52 micron) than those for C-protein. All soleus myofibrils label with anti-X but the centre-to-centre spacing was greater (0.67 micron). With plantaris myofibrils, where labelling occurs with anti-C or anti-H, the spacings resemble those in psoas myofibrils, but with anti-X the spacing resembles that in soleus myofibrils. The spacing of the fluorescent zones in an A-band, whether produced by anti-C, anti-X or anti-H does not vary with sarcomere length. We conclude that X-protein and H-protein, like C-protein, are thick filament components. With both fibres and myofibrils, there is no simple relationship between the amount of X-protein and the amount of C-protein. Many fast intermediate fibres in psoas and plantaris muscle label as strongly with anti-C as do fast white fibres but also label as strongly with anti-X as do fast and slow red fibres.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1983

H-protein and X-protein. Two new components of the thick filaments of vertebrate skeletal muscle.

With a view to obtaining a more complete view of the composition and structure of the thick filaments of vertebrate skeletal muscle, we have isolated and characterized two new myofibrillar components, H-protein and X-protein. These were purified by hydroxyapatite column chromatography of an impure C-protein preparation itself made from impure myosin extracted from rabbit back and leg muscles. H-protein is the protein responsible for band H on sodium dodecyl sulphate/polyacrylamide gel electrophoresis of crude myosin. X-protein, although present in such preparations in significant quantities, was not detected previously since it is difficult to resolve from C-protein by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. Physical-chemical parameters have been determined for the new proteins and compared with those of C-protein. The apparent chain weight of H-protein estimated by sodium dodecyl sulphate/polyacrylamide gel electrophoresis is 69,000, whereas that of X-protein (152,000) is only slightly greater than that of C-protein (140,000). The molecular weights of H- and X-proteins determined by sedimentation equilibrium centrifugation show that the molecules contain only a single polypeptide chain. The circular dichroism spectra indicate that the proteins have low alpha-helical contents. Both proteins, particularly H-protein, have a high proline content. Although X-protein is of similar chain weight to C-protein, the two show distinct differences in other properties. The sedimentation coefficient of X-protein is markedly lower than that of C-protein, suggesting X-protein is a more asymmetrical molecule. The amino acid compositions, although broadly similar, also show clear differences. Antibodies to H-protein, X-protein and C-protein have been raised in goats and shown not to cross-react.

Amino Acids↗
PubMed1978

The interaction of C-protein with heavy meromyosin and subfragment-2.

C-protein has previously been shown to bind to the light-meromyosin region of the myosin tail. Examination of mixtures of C-protein with heavy meromyosin or subfragment-2 or subfragment-1 in the analytical ultracentrifuge shows that there is also a binding site for C-protein in the subfragment-2 region of the tail.

Binding Sites↗
PubMedDate not supplied

A comparison of accuracy and estimated cost of methods for home blood glucose monitoring.

Venous serum glucose concentrations determined by a laboratory hexokinase technique were compared over a wide range of glucose concentrations with concentrations of capillary blood glucose determined by three reflectance meter techniques currently available in the United States (Eyetone and Dextrometer, Ames Company; StatTek, Bio-Dynamics BMC) and by visual interpretation of reagent strips (Chemstrip bG, Bio-Dynamics BMC). The Chemstrip bG reagent strip was read by patients, nurses, and a physician. In all cases, there was an excellent correlation between laboratory serum glucose concentrations and reflectance meter blood glucose determinations (r = 0.90-0.94, P less than 0.0001) or visual interpretation of Chemstrip bG (r = 0.85-0.92, P less than 0.0001). Chemstrip bG appears to be the least expensive method of glucose measurement. This method offers additional advantages in not requiring a reflectance meter, which needs frequent recalibration and other ancillary equipment for blood glucose determination.

Blood Glucose↗
PubMedDate not supplied

Characterization of a monoclonal antibody with CD44 like reactivity.

QE7.3E8 is a monoclonal antibody which precipitates two bands (90 and 160 kd) from a B cell line and four (90, 130, 146, and 160 kd) from monocytes. Whilst this immunoprecipitation pattern suggests that QE7.3E8 is a CD18 antibody, a number of other results are inconsistent with this interpretation. In particular, QE7.3E8 stains lymphoid tissue with a distribution clearly distinguishable from CD18 antibodies; it precipitates four bands from a T cell line which does not express the CD11b and CD11c alpha chains, and cross immunoprecipitation with QE7.3E8 and a CD11a antibody show that the molecules recognized by these antibodies are not associated. However, in tissue distribution and sequential precipitation experiments QE7.3E8 behaves like a CD44 antibody. CD44 identifies a single 85 kd protein, but high molecular weight complexes have been described, accounting for the multiple bands seen in immunoprecipitation studies. Sequential immunoprecipitation using QE7.3E8 and a Workshop-clustered monoclonal antibody (SBU 25-32) indicate that the QE7.3E8 reacts with the CD44 antigen.

Antibodies, Monoclonal↗
PubMedDate not supplied

The LFA-1 antigen in human B lymphocyte activation.

Human tonsil B cells include a subpopulation (30 per cent) of cells which lack LFA-1 antigen. Activation of tonsil B cells by culture with anti-IgM and interleukin-4 led to an increase in staining intensities and in the proportion of cells staining, until by 48 h the majority of B cells were positive. Culture of activated cells with low-molecular weight B cell growth factor, which induces a proportion of cells to proliferate, led to a minor further increase in expression of the LFA-1 antigen. Inclusion of a monoclonal antibody against the LFA-1 beta chain in culture did not affect either proliferation or immunoglobulin secretion. The expression of LFA-1 by B cells thus changes as B cells are activated, perhaps reflecting the changing requirements of B cells for interaction with other cells and tissue components. On the other hand, our results did not provide any support for the idea that the LFA-1 antigen is directly involved in the interaction of B cells with lymphokines which control proliferation and differentiation.

Antibodies, Monoclonal↗