Osmoregulation inBacillus subtilisunder potassium limitation: a new inducible K+-stimulated, VO43–-inhibited ATPase

2001 ◽  
Vol 47 (12) ◽  
pp. 1116-1125
Author(s):  
Jirí Šebestian ◽  
Zdenka Petrmichlová ◽  
Štepánka Šebestianová ◽  
Josef Náprstek ◽  
Jaroslava Svobodová
Keyword(s):  
Bacillus Subtilis ◽  
Atpase Activity ◽  
Synthetic Medium ◽  
Maximum Velocity ◽  
Monovalent Cation ◽  
Cross Reactivity ◽  
Potassium Transport ◽  
Cell Protein ◽  
P Type ◽  
Potassium Limitation

Bacillus subtilis exhibited an inducible K+-transporting ATPase activity with apparent Kmand maximum velocity Vmaxof 12.9 µM and 25.1 µmol·min–1·(g cell protein)–1, respectively, when cultivated on a synthetic medium containing less than 400 µM K+. Due to this enzyme, the growth rate of the bacterium in synthetic medium was not changed down to 115 µM K+, and the bacterium was able to grow down to 20 µM K+. The limiting K+concentration was higher in media with osmolarity increased by NaCl or sucrose. The ATPase was inhibited by micromolar concentrations of vanadate (Ki= 1.6 µM). The ATPase activity was not stimulated by any other monovalent cation. The subunit of this ATPase, with an Mrof 52 000, covalently bound the gamma phosphate group of ATP. This phosphorylated intermediate was unstable in neutral and basic pH as well as in the presence of potassium and was stable in acid pH. The enzyme did not show immunological cross-reactivity with antibody against Kdp ATPase of Escherichia coli.Key words: Kdp-like, potassium transport, Bacillus subtilis, transport ATPase, P-type ATPase.

Alterations in diaphragm contractility after nandrolone administration: an analysis of potential mechanisms

1999 ◽  
Vol 86 (3) ◽  
pp. 985-992 ◽  
Author(s):  
Michael I. Lewis ◽  
Mario Fournier ◽  
Amelia Y. Yeh ◽  
Paul E. Micevych ◽  
Gary C. Sieck
Keyword(s):  
Atpase Activity ◽  
Maximum Velocity ◽  
Interstitial Space ◽  
Myosin Atpase ◽  
Fiber Types ◽  
Intact Male ◽  
Cross Sectional ◽  
Myosin Atpase Activity ◽  
Relative Contribution

The aim of this study was to evaluate the potential mechanisms underlying the improved contractility of the diaphragm (Dia) in adult intact male hamsters after nandrolone (Nan) administration, given subcutaneously over 4 wk via a controlled-release capsule (initial dose: 4.5 mg ⋅ kg−1 ⋅ day−1; with weight gain, final dose: 2.7 mg ⋅ kg−1 ⋅ day−1). Control (Ctl) animals received blank capsules. Isometric contractile properties of the Dia were determined in vitro after 4 wk. The maximum velocity of unloaded shortening ( V o) was determined in vitro by means of the slack test. Dia fibers were classified histochemically on the basis of myofibrillar ATPase staining and fiber cross-sectional area (CSA), and the relative interstitial space was quantitated. Ca2+-activated myosin ATPase activity was determined by quantitative histochemistry in individual diaphragm fibers. Myosin heavy chain (MHC) isoforms were identified electrophoretically, and their proportions were determined by using scanning densitometry. Peak twitch and tetanic forces, as well as V o, were significantly greater in Nan animals compared with Ctl. The proportion of type IIa Dia fibers was significantly increased in Nan animals. Nan increased the CSA of all fiber types (26–47%), whereas the relative interstitial space decreased. The relative contribution of fiber types to total costal Dia area was preserved between the groups. Proportions of MHC isoforms were similar between the groups. There was a tendency for increased expression of MHC2B with Nan. Ca2+-activated myosin ATPase activity was increased 35–39% in all fiber types in Nan animals. We conclude that, after Nan administration, the increase in Dia specific force results from the relatively greater Dia CSA occupied by hypertrophied muscle fibers, whereas the increased ATPase activity promotes a higher rate of cross-bridge turnover and thus increased V o. We speculate that Nan in supraphysiological doses have the potential to offset or ameliorate conditions associated with enhanced proteolysis and disordered protein turnover.

Comparative force-velocity relation and analyses of myosin of dog atria and ventricles

1982 ◽  
Vol 243 (3) ◽  
pp. H391-H397 ◽  
Author(s):  
J. Wikman-Coffelt ◽  
H. Refsum ◽  
G. Hollosi ◽  
L. Rouleau ◽  
L. Chuck ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Calcium Concentration ◽  
Maximum Velocity ◽  
Calcium Sensitivity ◽  
Myofibrillar Proteins ◽  
Maximal Velocity ◽  
Velocity Of Shortening ◽  
Force Velocity ◽  
Normal Calcium ◽  
Actin Concentration

The isolated muscle and purified myofibrillar proteins of canine atria and ventricles were compared relative to force-velocity relations and rate of adenosine 5'-triphosphatase (ATPase) activity as a function of calcium concentrations. The maximal stress development of isolated trabeculae of canine atria was similar to that of canine right ventricular papillary muscles when analyzed at saturating calcium concentrations (7.5 mM); however, stress was less in the atria when studied at normal calcium concentrations (2.5 mM). The maximal velocity of shortening of atrial trabeculae was about 2.3 times higher than that of ventricular muscle. Regulated actomyosin characterized from the myofibrillar proteins of the two tissues gave directionally similar calcium sensitivity. The maximum velocity of shortening for actin-activated atrial myosin of the dog was approximately 1.8 times higher when the latter was analyzed as a function of actin concentration. Both maximal tension of isolated muscle and regulated actomyosin ATPase activity were dependent on calcium concentration.

Evaluation of an alternative S100b assay for use in cardiac surgery: relationship with microemboli and neuropsychological outcome

Perfusion ◽  
2007 ◽  
Vol 22 (4) ◽  
pp. 267-272 ◽  
Author(s):  
D.C. Whitaker ◽  
A.J.E. Green ◽  
J. Stygall ◽  
M.J.G. Harrison ◽  
S.P. Newman
Keyword(s):  
Cardiac Surgery ◽  
Limit Of Detection ◽  
Artery Bypass ◽  
Sandwich Elisa ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Cell Protein ◽  
Skin Closure ◽  
Neuropsychological Outcome ◽  
The Right

Introduction. The aim of the study was to investigate the relationship between S100b release, neuropsychological outcome and cerebral microemboli. Peri-operative assay of the astroglial cell protein S100b has been used as a marker of cerebral damage after cardiac surgery but potential assay cross-reactivity has limited its specificity. The present study uses an alternative enzyme-linked immunoabsorbant assay (ELISA) for serum S100b that has documented sensitivity and specificity data in patients undergoing coronary artery bypass grafting (CABG). Methods. Fifty-five consecutive patients undergoing routine CABG surgery received serial venous S100b sampling at five time points: i) Pre-operative, ii) At the end of cardiopulmonary bypass (CPB), iii) 6 hrs, iv) 24 hrs and v) 48 hrs post skin closure. A previously described sandwich ELISA with monoclonal anti- S100b was used. This assay has a lower limit of detection of 0.04 μ g/L and < 0.006% reactivity with S100a at a concentration of 100 μg/L S100a. Cerebral microemboli during surgery were recorded by transcranial Doppler monitor over the right middle cerebral artery. Evidence of cerebral impairment was obtained by comparing patients' performance in a neuropsychological battery of 9 tests administered 6—8 weeks post-operatively with their pre-operative scores. Results. There was a significant increase in S100b only at the end of bypass (mean 0.30 μg/L, SD ± 0.33 and range .00 to 1.57). S100b levels at the end of bypass did not correlate with neuropsychological outcome or microemboli counts. Conclusions. The low levels of S100b detected using the present assay, despite its high sensitivity and despite the routine use of cardiotomy suction, suggest that the assay may have higher specificity for cerebral S100b than previously used assays. There was no evidence that this assay is related to neuropsychological change or cerebral microemboli in cardiac surgery. Perfusion (2007) 22, 267—272.

Regulation of renal Na+-K+-ATPase in the rat: role of increased potassium transport

1986 ◽  
Vol 251 (2) ◽  
pp. F199-F207
Author(s):  
S. K. Mujais ◽  
M. A. Chekal ◽  
J. P. Hayslett ◽  
A. I. Katz
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Potassium Transport ◽  
High Potassium ◽  
Dietary Potassium ◽  
Collecting Tubule ◽  
Physiological Dose ◽  
Potassium Loading ◽  
Cortical Collecting Tubule

The purpose of this study was to characterize the alterations in collecting tubule Na+-K+-ATPase activity produced by sustained increments in dietary potassium in the rat and to evaluate the role of aldosterone in their generation. In adrenal-intact animals, feeding a high-potassium diet (10-fold that of control) or administration of a high physiological dose of aldosterone (5 micrograms X 100 g-1 X day-1), which simulates the delivery rate of this hormone during potassium loading (both for 7 days), caused marked increments in Na+-K+-ATPase activity in the cortical collecting tubule (CCT) but had no effect on the enzyme in the inner stripe of the medullary collecting tubule (MCT). A significant increase in enzyme activity was also observed after smaller dietary potassium increments (2.5 and 5 times the control) and after 4 (but not 2) days of dietary potassium load. In adrenalectomized rats provided with physiological replacement doses of corticosterone and aldosterone (0.8 micrograms X 100 g-1 X day-1), Na+-K+-ATPase activity in both CCT and MCT was similar to that of adrenal-intact controls but remained unchanged after 7 days on the potassium-enriched (10-fold) diet. In contrast, adrenalectomized animals receiving the high physiological dose of aldosterone displayed an increase in Na+-K+-ATPase activity of CCT comparable with that of adrenal-intact animals, whereas the enzyme activity in the MCT was unaffected. In conclusion, 1) following chronic potassium loading Na+-K+-ATPase activity increases significantly in the CCT with no change in its activity in the inner stripe of the MCT.(ABSTRACT TRUNCATED AT 250 WORDS)

NH2-terminal-inserted myosin II heavy chain is expressed in smooth muscle of small muscular arteries

1997 ◽  
Vol 272 (5) ◽  
pp. C1532-C1542 ◽  
Author(s):  
M. E. DiSanto ◽  
R. H. Cox ◽  
Z. Wang ◽  
S. Chacko
Keyword(s):  
Smooth Muscle ◽  
Atpase Activity ◽  
Heavy Chain ◽  
Light Chain ◽  
Maximum Velocity ◽  
Myosin Isoforms ◽  
Coding Region ◽  
Shortening Velocity ◽  
Visceral Smooth Muscle ◽  
Saphenous Artery

We demonstrate, using reverse transcriptase-polymerase chain reaction, that, whereas abdominal aorta from rabbit consists almost entirely of myosin heavy chain (MHC) mRNA with no insert at the 5'-terminal coding region, the distributing arteries (femoral and saphenous) begin to show MHC mRNA with the 21-nucleotide insert that encodes seven amino acids in the ATP-binding region located in the myosin head. The femoral/iliac artery contains > 50% inserted mRNA, whereas the more distal saphenous artery contains > 80% inserted mRNA. This insert is also present in the smooth muscle from rat tail artery but is absent in the smooth muscle from rat aorta. The actin-activated ATPase activity of myosin from the rabbit femoral/saphenous artery is 1.7-fold higher than that of the myosin from the aorta. A concomitant increase (about twofold) in the maximum shortening velocity of the saphenous artery, compared with that of the aorta, indicates that the preponderance of the inserted myosin is associated with both an increase in the actin-activated ATPase activity and a larger maximum velocity of shortening. Furthermore, analysis of the 17-kDa essential light chain from the aorta reveals near equal quantities of the 17-kDa light chain isoforms a and b, whereas the myosin from the femoral/ saphenous artery contains predominantly the 17-kDa light chain a isoform. Together, these data indicate that the smooth muscle cells from the small distributing arteries are similar to those of visceral smooth muscle with respect to the expression of myosin isoforms, actin-activated myosin ATPase activity and contractility.

Activated neutrophils inhibit Na(+)-K(+)-ATPase in canine renal basolateral membrane

1992 ◽  
Vol 262 (6) ◽  
pp. C1364-C1370 ◽  
Author(s):  
C. M. Julin ◽  
J. J. Zimmerman ◽  
V. Sundaram ◽  
M. C. Chobanian
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Hypochlorous Acid ◽  
Basolateral Membrane ◽  
Maximum Velocity ◽  
Reactive Oxygen Metabolites ◽  
Activated Neutrophils ◽  
Hydroxyl Radical Scavengers ◽  
Atpase Inhibition ◽  
Oxygen Metabolites

To examine the effects of activated neutrophils (PMNs) on Na(+)-K(+)-ATPase, phorbol 12-myristate 13-acetate (PMA)-stimulated PMNs were incubated with canine renal cortical basolateral membrane (BLM), and BLM ouabain-sensitive Na(+)-K(+)-ATPase activity was subsequently quantified. Na(+)-K(+)-ATPase activity decreased to 40.0 +/- 8.7% (SE) of control in the presence of activated PMNs, from 0.89 +/- 0.12 to 0.34 +/- 0.05 mumol Pi.mg protein-1.min-1. This inhibition coincided with a decrease in the apparent Michaelis constant (Km) for ATP from 0.18 +/- 0.02 to 0.05 +/- 0.01 mM. Inclusion of catalase (CAT) and superoxide dismutase (SOD) in the BLM/PMN/PMA incubation mixture resulted in partial preservation of enzyme activity, with an increase to 57.0 +/- 4.6% of control with CAT alone and to 70.0 +/- 5.3% with both CAT and SOD. SOD alone had no protective effect. Neither the myeloperoxidase inhibitor azide nor the hypochlorous acid scavenger L-methionine preserved enzyme activity. Hydroxyl radical scavengers and iron chelators were also ineffective in attenuating Na(+)-K(+)-ATPase inhibition by activated PMNs. These results indicate that activated PMNs mediate a decrease in BLM Na(+)-K(+)-ATPase activity characterized by a reduction in maximum velocity and Km for ATP that appears to be mediated in part by reactive oxygen metabolites.

Sign in / Sign up

Export Citation Format

Share Document