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J D Potter

Publications and source records attributed to J D Potter.

At least 235 records · Page 13Linked to original sources
PubMed1990

Rapid activation by photolysis of nitr-5 in skinned fibres of the striated adductor muscle from the scallop.

Photolysis of nitr-5, a caged calcium molecule, has been used for rapid activation of skinned fibre bundles of a myosin-regulated muscle, the striated adductor of the scallop, Pecten maximus. Chemically skinned fibre bundles (diameter 70-200 microns) were equilibrated in solutions containing 1-3 mM nitr-5 (pCa 6.1) and then activated by ultraviolet laser pulse (25 ns). Pulse energies of 60-95 mJ gave contractions of over 90% maximum tension and a mean half-time for tension rise of 43 ms (n = 4) at 12 degrees C. Electrically stimulated bundles of intact fibres develop a tetanus with a rise half-time of 60.2 ms at 10 degrees C (n = 5) (Rall, J.A. (1981) J. Physiol. 321, 287-295, and personal communication). At lower pulse energies the skinned fibres gave smaller amplitude contractions with slower rates of rise (up to 260 ms half-time). In addition, a slower component of tension development (mean rise half-time 13.3 s) was often observed. In ATP-free solutions containing hexokinase and glucose, rigour tension developed with a delayed onset. Rapid release of ATP (0.47-0.59 mM) from photolysis of caged ATP (2 mM) at pCa 4.5 then caused a rapid contraction with a mean half-time for tension development of 17 ms (n = 4). The fast activation rates obtained by the photorelease of Ca2+ from nitr-5 are similar to those obtained with skinned skeletal fibres of actin-regulated muscle. The results imply that the rate-limiting step in excitation-contraction coupling of the scallop muscle is not the increase in sarcoplasmic Ca2+, but rather the activation of the muscle in response to this increase. The half-times of ATP-induced contractions at pCa 4.5 suggest that in a contraction activated by a rapid Ca2+ jump the process comprising ATP hydrolysis and cross-bridge cycling occurs at a somewhat faster rate than the Ca2(+)-dependent activation process which precedes it.

Adenosine Triphosphate↗
PubMed1990

Mechanical characteristics of skinned and intact muscle fibres from the giant barnacle, Balanus nubilus.

Intact muscle fibres from Balanus nubilus develop tensions of up to 600 kN.m-2 during electrical stimulation. The rise of tension occurs with a half-time (177 ms at 12 degrees C) about fivefold longer than that of tetanised frog muscle at the same temperature. The response of myofibrillar bundles to a rapid stretch resembles that of frog muscle but has a yo value (i.e. the size of an instantaneous release necessary to just discharge tension) which is ca. 2.5 times smaller, and phase 2 of the tension transient (the "quick phase") occurs at a rate comparable to that of frog muscle. In contrast, the ATPase activity (0.018 mmoles.kg wet weight-1.s-1) of this preparation and its maximum shortening velocity (0.15-0.16 muscle lengths.s-1) are both at least fivefold slower than frog muscle. These findings can be accounted for by a cross-bridge cycle in barnacle muscle in which events prior and subsequent to the tension generating step(s) occur at a rate at least fivefold slower than comparable steps in frog muscle, but the step(s) associated with tension development occur at similar rates in the two preparations. Since the rate of mechanical relaxation in barnacle muscle is modified in the presence of intracellular calcium buffers and by depolarisation-induced elevation of the free calcium during the relaxation phase, it is proposed that the time course of relaxation is not determined exclusively by the kinetics of the cross-bridge cycle, but is also dependent on the free calcium concentration during relaxation.

Adenosine Triphosphatases↗
PubMed1990

Increased incidence of carcinoma of the breast associated with abdominal adiposity in postmenopausal women.

Epidemiologic studies have established that increased body weight is associated with a greater incidence of breast cancer in postmenopausal women. The authors hypothesized that abdominal adiposity further increases risk of postmenopausal breast cancer. They therefore investigated the waist-to-hip circumference ratio in relation to breast cancer incidence in a nested case-control study of 41,837 postmenopausal Iowa women aged 55-69 years. Women were recruited through a mail survey in January 1986 and were asked to have someone measure their body circumferences using a paper tape measure and written instructions. Cancer incidence was ascertained using a statewide cancer registry. A total of 229 incident breast cancers occurred among at-risk women during the first 2 years of follow-up (1986-1987). Compared with randomly selected controls (n = 1,839), women with incident breast cancer had a higher age-adjusted mean waist-to-hip ratio (by 0.013 units, p = 0.030), as well as greater mean weight (by 1.7 kg, p = 0.07), body mass index (by 0.6 kg/m2, p = 0.08), and weight gain since age 18 (by 2.7 kg, p less than 0.01). In multiple logistic regression models, age and current body mass index were significant effect modifiers of the association between the waist-to-hip ratio and breast cancer. A two-standard deviation increase in the waist-to-hip ratio (0.168 units) was associated with no increase in the relative risk of breast cancer in younger and lighter postmenopausal women. However, in older, heavier postmenopausal women, the same increase in the waist-to-hip ratio carried greater than a twofold excess relative risk. Adjustment for other breast cancer risk factors did not materially alter this finding. Two plausible explanations for elevated breast cancer incidence in women with abdominal adiposity include 1) increased concentrations of non-protein-bound estrogens due to reduced sex hormone binding globulin, or 2) increased conversion of adrenal androgens to estrone with abdominal adiposity.

Abdominal Muscles↗
PubMed1990

The association of body fat distribution with lifestyle and reproductive factors in a population study of postmenopausal women.

We examined the cross-sectional association of fat distribution with a number of lifestyle and reproductive factors in a random sample of 40,980 postmenopausal women, aged 55 to 69 years. The relationship of weight history with current fat distribution was also explored. Body fat distribution, defined by the ratio of waist-to-hip circumferences (WHR), was most strongly related to body mass index (BMI). WHR was also significantly and negatively associated with physical activity, alcohol consumption, and education, and was significantly positively associated with age, cigarette smoking, and a number of reproductive factors, such as history of infertility, number of live births, age at first live birth, and replacement estrogen use. WHR was not related to past BMIs, after adjusting for current body mass. The best-fit final regression model included (beta +/- s.e.) age (0.003 +/- 0.0001), BMI (0.02 +/- 0.0005), a curvilinear term for BMI (-0.0002 +/- 0.00001), greater than high school education (-0.007 +/- 0.001), cigarette smoking (0.02 +/- 0.001), alcohol drinker (-0.005 +/- 0.001), and number of live births (0.001 +/- 0.0002). The model R2 was 0.21. BMI (R2 = 0.18) and age (R2 = 0.02) accounted for most of the variation in WHR. The association of these lifestyle and reproductive factors with WHR suggests that the deposition of fat in the abdominal versus the gluteal region may be influenced somewhat by factors other than overall adiposity and genetics.

Abdomen↗
PubMed1989

Association of incident carcinoma of the endometrium with body weight and fat distribution in older women: early findings of the Iowa Women's Health Study.

Previous epidemiological studies have demonstrated that obesity increases endometrial cancer risk two- to 10-fold. To test the hypothesis that abdominal adiposity further increases this relative risk, we conducted a nested case-control study of endometrial cancer incidence in a cohort of 41,873 women ages 55-69 years. Women were recruited by mail and asked to have a friend measure circumferences of several body parts using a tape measure and written instructions. Two-year follow-up for cancer incidence was conducted using a state-wide cancer registry. Compared to random controls (n = 1,274), cases (n = 63) had higher age-adjusted mean values of waist-to-hip circumference ratio (P = 0.10) and trunk-to-limb circumference ratio (waist plus hip circumferences divided by arm plus calf circumferences, P = 0.008). Other anthropometric variables, including current body mass index and current weight, were also greater (P less than 0.001) in cases than controls. After accounting for the association with body mass index, neither the waist-to-hip ratio nor the trunk-to-limb ratio remained associated with endometrial cancer incidence (P greater than 0.40). A 5 kg/m2 increase in body mass index was associated with an adjusted relative risk of endometrial cancer of 1.80 [95% CI = 1.46, 2.22] when other significant risk factors, namely age, education level, extended use of exogenous estrogens, and age at menopause, were taken into account. We conclude that endometrial cancer risk is increased in relation to the amount but not the distribution of adiposity. This is in contrast with several other diseases in which, in addition to overall body mass, the distribution of adiposity is also important.

Abdomen↗
PubMed1989

Reciprocal coupling between troponin C and myosin crossbridge attachment.

The attachment of cycling myosin crossbridges to actin and the resultant muscle contraction are regulated in skeletal muscle by the binding of Ca2+ to the amino-terminal, regulatory sites of the troponin C (TnC) subunit of the thin filament protein troponin. Conversely, the attachment of crossbridges to actin has been shown to alter the affinity of TnC for Ca2+. In this study, fluorescently labeled TnC incorporated into reconstituted thin filaments was used to investigate the relationship between crossbridge attachment to actin and structural changes in the amino-terminal region of TnC. Fluorescence intensity changes were measured under the following conditions: saturating [Ca2+] in the absence of crossbridges, rigor crossbridge attachment in the presence and absence of Ca2+, and cycling crossbridge attachment. The percent of heavy meromyosin crossbridges associated with the thin filaments under these conditions was also determined. The results show that, in addition to the binding of Ca2+ to TnC, the attachment of both rigor and cycling crossbridges to actin alters the structure of TnC near the regulatory, Ca2+-specific sites of the molecule. A differential coupling between weakly versus strongly bound crossbridge states and TnC structure was detected, suggesting a possible differential regulation of these states by conformational changes in TnC. These findings illustrate a reciprocal coupling, via thin filament protein interactions, between structural changes in TnC and the attachment of myosin crossbridges to actin, such that each can influence the other, and indicate that TnC is not simply an on-off switch but may exist in a number of different conformations.

Actins↗
PubMed1988

Cadmium-substituted skeletal troponin C metal binding investigations and sequence assignment of the cadmium-113 resonances.

The binding of cadmium to the calcium binding subunit of skeletal troponin (STnC) has been reinvestigated using direct binding methods and fluorescent derivatives. These data provide straightforward explanations of the observed titration behavior in the 113Cd NMR (Ellis, P.D., Strang, P., and Potter, J.D. (1984) J. Biol. Chem. 259, 10348-10356). Further, fluorescent derivatives of skeletal troponin C provide an excellent means of establishing a sequence assignment for the resonances observed in the 113Cd NMR. The results of these experiments demonstrate that sites I and II, the Ca2+ regulatory sites, can be assigned to resonances at -108.5 and -101.5 ppm, respectively. Sites III and IV, the structural sites, are assigned to resonances -112.8 and -106.8 ppm, respectively. These data are discussed in terms of recent structural findings and speculations.

Animals↗
PubMed1988

Amino acid sequence of bovine cardiac troponin I.

Troponin I (TnI) is the inhibitory subunit of troponin, the thin filament regulatory complex which confers calcium sensitivity to striated muscle actomyosin ATPase activity. We have determined the amino acid sequence of TnI from adult bovine cardiac muscle. This protein is a single polypeptide chain of 211 amino acids with an acetylated amino terminus, a calculated molecular weight of 23,975, and a net charge of +17 at neutral pH. There was no evidence for heterogeneity of the sequence. Comparison with other available TnI sequences shows an amino-terminal extension of 27-33 residues which is present in cardiac but not skeletal TnI. The remainder of the polypeptide is common to both cardiac and skeletal TnI. In the amino-terminal half of the common polypeptide, only 29% of the residues are invariant in all sequences. The carboxyl-terminal half (residues 124-210) is much more highly conserved, with 66% invariant residues. Bovine cardiac TnI and rabbit cardiac TnI are very similar in sequence: only 12 of 26 residues are identical in the amino-terminal segments, but the remaining residues of the proteins are 97% identical.

Amino Acid Sequence↗
PubMed1988

Skinned ventricular fibres: troponin C extraction is species-dependent and its replacement with skeletal troponin C changes Sr2+ activation properties.

Troponin C (TnC) was extracted from bundles of rat and mouse skinned cardiac ventricular cells by a method similar to that previously used to extract TnC from skinned rabbit skeletal muscle cells (Kerrick et al., J. biol. Chem. 260, 15687-93, 1985) and replaced with either bovine cardiac or rabbit fast-twitch skeletal TnC. In contrast, the same TnC extraction conditions will not extract TnC from either bovine or rabbit cardiac skinned fibres. When the extracted TnC was replaced by bovine cardiac TnC the Sr2+-activated tension relationship was not altered from control values. In contrast, replacement of the endogenous TnC with exogenous rabbit fast-twitch TnC caused the relationship between Sr2+ concentration and tension to shift towards higher concentrations of Sr2+. The Sr2+-activated tension of rat fibres with fast-twitch TnC was identical to that of rabbit fast-twitch skinned fibres. Partial skeletal TnC substitution in skinned cardiac cells gradually shifted the relationship between tension and [Sr2+] to higher Sr2+ concentrations and caused the cells to be activated over a wider range of Sr2+ concentrations. Thus it appears that the activation of rat cardiac skinned cells by Sr2+ is determined by characteristics of the TnC. In contrast, the Sr2+ activation of skinned rabbit fast-twitch skeletal fibres containing either cardiac or skeletal TnC is identical, strongly suggesting that protein-protein interactions determined the Sr2+ -activation properties in these fibres.

Animals↗
PubMed1988

Transient kinetics and time-resolved X-ray diffraction studies in isolated single muscle fibres.

The timing of events associated with the contraction and relaxation of the force cycle is described in isolated single arthropod muscle fibers using the fluorescently labelled derivatives of the Ca2+ binding sub-unit of troponin TnC. The kinetics of the subtracted fluorescence (490-410 nm) response from injected TnCDANZ, labelled at the Ca2+ specific sites, shows a rapid rise which is some 90% complete at 50% force consistent with rapid Ca2+ binding to this sub-unit. Subsequently the TnCDANZ fluorescence decays 2x more slowly, at 12 degrees C, than force consistent with a slower release of this bound Ca2+. In fibers injected with both aequorin and TnCDANZ, the aequorin kinetics are essentially unaltered compared to control fibers in the presence of 10-100 microM TnCDANZ. The peak of the aequorin response occurs some 150-170 msec in front of the TnCDANZ peak and the T 1/2 for light decay is faster than either force or TnCDANZ decay, but there is a 'tail' to the aequorin light response (elevated free Ca2+) well into the relaxation phase, seen both in cannulated and intact muscle fibers. The kinetics of the fluorescence of TnCIAANS, labelled of the Ca2+-Mg2+ sites, shows a slow decrease (T 1/2 1.8 sec) and subsequent increase (T 1/2 2.5 sec) in fluorescence consistent with a slow loading and unloading of these sites with Ca2+ during a tetanus. Time resolved X-ray diffraction from intact muscle fibers indicate that forces of up to 600 kN/m2 can be developed at sarcomere lengths of 8-10 micron. Force shows a marked sarcomere dependency while the aequorin response is relatively insensitive. At these high forces, there is a marked change in intensity of the first actin layer line (A2 at 38 nm), consistent with S1 (cross-bridge) attachment, which has a T 1/2 for rise of 125-150 msec.

Actins↗
PubMed1987

Isolation and sequence of a cDNA clone for rabbit fast skeletal muscle troponin C. Homology with calmodulin and parvalbumin.

The binding of Ca2+ to troponin C (TnC) regulates skeletal muscle contraction. We have isolated a full-length cDNA clone for fast skeletal muscle TnC from a neonatal rabbit skeletal muscle library and determined its nucleic acid sequence. The amino acid sequence deduced from this clone matches the previously reported amino acid sequence (Collins, J. H., Greaser, M. L., Potter, J. D., and Horn, M. J. (1977) J. Biol. Chem. 252, 6356-6362) except at the amino terminus. According to the nucleotide sequence, the first 2 residues of TnC are threonine-aspartic acid, which is the reverse of the order reported previously. The isolation of the adult form of TnC from a neonatal library suggests that there may be no developmental isoforms of fast TnC. The protein coding region of the fast TnC clone has 67% homology with the reported nucleotide sequence for chicken slow TnC (Putkey, J. A., Carroll, S. L., and Means, A. R. (1987) Mol. Cell. Biol. 7, 549-1553). The homologies between the nucleotide sequences of TnC, calmodulin, and parvalbumin provide evidence that all three proteins were derived from a common precursor molecule which had four Ca2+-binding sites.

Animals↗
PubMed1987

Bovine cardiac troponin T: amino acid sequences of the two isoforms.

Troponin T (TnT) is the tropomyosin-binding subunit of troponin, the thin filament regulatory complex that confers calcium sensitivity to striated muscle contraction and actomyosin ATPase activity. Bovine cardiac muscle contains two isoforms (TnT-1 and TnT-2) of TnT that differ in sequence near their amino termini. Thin filaments containing TnT-2 require less calcium to activate the MgATPase rate of myosin than do thin filaments containing TnT-1. Using whole troponin T purified from adult bovine cardiac muscle, we have determined the complete amino acid sequence of the larger, more abundant isoform TnT-1. We confirmed that sequence differences between TnT-1 and TnT-2 are confined to the amino-terminal regions and found that TnT-1 makes up approximately 75% of the total troponin T isolated. Partial sequencing of the separated isoforms showed that the difference between them is due solely to residues 15-19 (Glu-Ala-Ala-Glu-Glu) of TnT-1 being absent from TnT-2. The deleted segment may correspond to the product of exon 4 of the chicken cardiac TnT gene [Cooper, T.A., & Ordahl, C.P. (1985) J. Biol. Chem. 260, 11140-11148]. Exon 5, which is developmentally regulated in the chicken, is not expressed in either TnT-1 or TnT-2. TnT-1 contains 284 amino acid residues and has a Mr of 33,808, while TnT-2 contains 279 amino acid residues and has a Mr of 33,279. Bovine cardiac TnT contains the only known thiol group in any isolated TnT (Cys-39 of TnT-1, Cys-34 of TnT-2). Comparison of bovine, rabbit, and chicken cardiac TnT sequences shows near identity of the amino-terminal 13 amino acid residues (exons 2 and 3 of the chicken cardiac gene), many differences in the following 60 residues (exons 4-8), and great similarity in the C-terminal 230 residues (exons 9-18).

Amino Acid Sequence↗
PubMed1987

Effect of rigor and cycling cross-bridges on the structure of troponin C and on the Ca2+ affinity of the Ca2+-specific regulatory sites in skinned rabbit psoas fibers.

Intrinsic troponin C (TnC) was extracted from small bundles of rabbit psoas fibers and replaced with TnC labeled with dansylaziridine (5-dimethylaminonaphthalene-1-sulfonyl). The flourescence of incorporated dansylaziridine-labeled TnC was enhanced by the binding of Ca2+ to the Ca2+-specific (regulatory) sites of TnC and was measured simultaneously with force (Zot, H.G., Güth, K., and Potter, J.D. (1986) J. Biol. Chem. 261, 15883-15890). Various myosin cross-bridge states also altered the fluorescence of dansylaziridine-labeled TnC in the filament, with cycling cross-bridges having a greater effect than rigor cross-bridges; and in both cases, there was an additional effect of Ca2+. The paired fluorescence and tension data were used to calculate the apparent Ca2+ affinity of the regulatory sites in the thin filament and were shown to increase at least 10-fold during muscle activation presumably due to the interaction of cycling cross-bridges with the thin filament. The cross-bridge state responsible for this enhanced Ca2+ affinity was shown to be the myosin-ADP state present only when cross-bridges are cycling. The steepness of the pCa force curves (where pCa represents the -log of the free Ca2+ concentration) obtained in the presence of ATP at short and long sarcomere lengths was the same, suggesting that cooperative interactions between adjacent troponin-tropomyosin units may spread along much of the actin filament when cross-bridges are attached to it. In contrast to the cycling cross-bridges, rigor bridges only increased the Ca2+ affinity of the regulatory sites 2-fold. Taken together, the results presented here indicate a strong coupling between the Ca2+ regulatory sites and cross-bridge interactions with the thin filament.

Animals↗
PubMed1987

The effect of [Mg2+] on the Ca2+ dependence of ATPase and tension development of fast skeletal muscle. The role of the Ca2+-specific sites of troponin C.

Conflicting reports have appeared concerning the effect of [Mg2+] on muscle activity. Several groups have found that increasing [Mg2+] produces a right-ward shift of the pCa-tension curve, while others have found no effect of [Mg2+] on myofibrillar ATPase activity. The present study is a careful evaluation of the effect of [Mg2+] on myofibrillar ATPase, skinned fiber tension development, TnCDANZ (troponin C (TnC)-labeled with 5-dimethylaminonaphthalene-1-sulfonyl aziridine) fluorescence, and simultaneous TnCDANZ fluorescence and tension development in the same fiber. A small effect of [Mg2+] on both ATPase and tension development was found with an apparent association constant of about 2 X 10(2) M-1. The Ca2+ dependence of TnCDANZ fluorescence was similarly effected by [Mg2+], either alone or when incorporated into TnC-depleted skinned fibers (K'Mg approximately equal to 2-3 X 10(2) M-1), suggesting that the effect of [Mg2+] on activity is due to an effect of [Mg2+] on Ca2+ binding to the Ca2+-specific sites of TnC. It is not yet clear whether this effect of [Mg2+] is through direct competition at the binding sites or through indirect effects. In either case, the calculated effect of physiological [Mg2+] is so small that the regulatory sites of TnC can still be considered "Ca2+-specific." In addition, a slightly greater effect of [Mg2+] on tension development (K'Mg = 4.62 X 10(2) M-1) was observed only for very low levels of [Mg2+], which might suggest an additional effect of Mg2+ on tension development which is saturated by millimolar Mg2+.

Animals↗
PubMed1987

Calcium binding and fluorescence measurements of dansylaziridine-labelled troponin C in reconstituted thin filaments.

The direct binding of Ca2+ to reconstituted thin filaments containing troponin C and the 5-dimethylaminonaphthalene-1-sulphonylaziridine (DANZ) fluorescent analogue of troponin C (TnCDANZ) was measured (25 degrees C) at three Mg2+ concentrations. Biphasic Scatchard plots were found for all binding curves reflecting the binding of Ca2+ to high- and low-affinity sites of troponin. The binding of Ca2+ to the high-affinity sites had a greater sensitivity to Mg2+ (KMg = 1 x 10(4)M-1) than the low-affinity sites (KMg = 1.2 x 10(3)M-1). The fluorescence change of thin filaments reconstituted with TnCDANZ was titrated with Ca2+ in the same solutions used for binding assays. The Ca2+-dependent fluorescence change had nearly the same sensitivity to Mg2+ (KMg = 9.4 x 10(2)M-1) as did Ca2+ binding to the low-affinity sites. The Ca2+ concentration at the midpoint of the fluorescence change was about 0.3 log units less than at the midpoint for Ca2+ binding to the low-affinity sites. A similar relationship between the fluorescence change and Ca2+ binding to the low-affinity sites of isolated TnCDANZ was measured (4 degrees C). From these results the binding of Ca2+ to either low-affinity site is concluded to produce the fluorescence change. In comparison with the low-affinity sites of isolated troponin and troponin-tropomyosin complex, the low-affinity sites of reconstituted thin filaments were consistently lower in Ca2+ affinity.

Actin Cytoskeleton↗