Permeability to cobalt ions and action of colicin K in a mutant that overproduces cardiolipin.

A mutant of Escherichia coli that produces excess cardiolipin becomes less capable of transporting Co2+. Cardiolipin therefore does not act as an ionophore under these conditions. Colicin K brings about the typical increase in permeability to Co2+ in the mutant.

Phospholipase activity plays no role in the action of colicin K.

1. A mutant lacking both detergent-resistant and detergent-sensitive phospholipase A activities is fully sensitive to colicin K. 2. In the absence of cellular phospholipases A, colicin K does not promote hydrolysis of phosphatidylethanolamine. 3. Cells of the colicin-treated mutant lacking lysophosphatidylethanolamine are as abnormally permeable to Co-2+ as the wind type is. 4. Increased levels of lysophosphatidylethanolamine in colicin-treated cells are not necessary for the increased sensitivity to sodium dodecylsulfate.

Enzyme reaction rates and the stochastic theory of kinetics.

The kinetics of an elementary reaction step are discussed from the viewpoint of the stochastic theory of chemical kinetics. The general form of the rate constant found in the stochastic approach is described and compared with the expression from transition state theory. Whereas the stochastic theory predicts a rate enhancement in cases which are not adiabatic (in the quantum mechanical sense), transition state theory, which is essentially an adiabatic theory of reaction rates, does not permit inclusion of the effect. This effect can be expected to be of greater importance in cases of catalysis by structures, such as enzymes, containing large numbers of vibrational degrees of freedom (particularly low frequency ones) than in cases lacking such structures. The stochastic theory is more general than the transition state theory, the rate constant expression given by the latter being obtainable from the former when restrictive assumptions, including that of adiabaticity, are made. Interpretations of enzyme catalysis based on the transition state theory must thus be viewed as speculative.

The incise drape.

Since it is impossible to sterilize the surgical field in ocular surgery, postoperative endophthalmitis remains a serious complication of intraocular surgery. The surgeon should take all reasonable precautions to avoid this disaster. We believe that this includes isolating the skin and eyelids from the operative field with the incise drape.

Altered phospholipid metabolism in a sodium-sensitive mutant of Escherichia coli.

A mutant of Escherichia coli has been isolated, the growth of which is inhibited by low concentrations (1 mm) of NaCl. High levels of magnesium, calcium, or strontium in the medium permit growth in the presence of sodium. The metal content of the inhibited mutant is normal, but the strain is unable to tolerate levels of sodium to which the wild type is indifferent. Immediately after the addition of sodium to cultures of the mutant, rates of synthesis of protein, ribonucleic acid, deoxyribonucleic acid, and total lipid are unchanged, but more cardiolipin and less phosphatidylethanolamine are produced. The direct enzymatic cause of this change, which affects membrane function, is not known. Studies of the metabolism of phosphatidylglycerol in vivo after pulse-labeling with [2-(3)H]glycerol reveal that a major pathway both in wild-type and mutant strains involves the cleavage of labeled glycerol from phosphatidylglycerol.

Effects of colicins E1 and K on permeability to magnesium and cobaltous ions.

The energy-dependent exchange of intracellular Mg(2+) with extracellular Mg(2+) or Co(2+) is inhibited by colicin E1 and, less strongly, by colicin K. Treatment with either colicin causes a net loss of intracellular Mg(2+). This loss begins immediately in cells treated with colicin E1, but in colicin K-treated cells the onset of Mg(2+) loss is delayed 1 to 10 min, depending upon the temperature and the multiplicity of colicin K. Both colicins differ from chemical inhibitors of energy-yielding metabolism; energy poisons block transport of Mg(2+) and Co(2+), but both colicins increase passive permeability to Mg(2+) and Co(2+). Inhibitors of energy-yielding metabolism (and of Mg(2+) exchange) block the initiation of Mg(2+) loss by either colicin, but do not stop colicin-promoted efflux once it has begun. Colicin E1 added before colicin K prevents the more rapid Mg(2+) efflux characteristic of colicin K-treated cells. Quantitative comparisons of the effects of colicins E1 and K upon permeability to Mg(2+) and Co(2+) lead us to conclude that the two colicins are not identical in their mode of action.

Disappearance of specific proteins from the membranes of colicin-treated cells.

Two colicins that affect energy metabolism in Escherichia coli (colicins K and E1) are shown to cause loss of specific membrane proteins from treated cells. Disappearance of these proteins after treatment with colicin K occurs at low multiplicities and is independent of ATPase (EC 3.6.1.4) and phospholipase A (EC 3.1.1.4) activities. The uncouplers carbonyl cyanide m-chlorophenylhydrazone and dinitrophenol do not alter the pattern of membrane proteins.

Viability of Escherichia coli treated with colicin K.

Conditions permitting survival (colony formation) of E. coli after treatment with colicin K have been found. Survival required K+ and Mg2+ at concentrations high enough to replace the intracellular ions lost from colicintreated cells. Either glucose minimal medium or broth could support survival. Survival was still observed after colicin-promoted efflux of Rb+ and decline in ATP levels had occurred, and after the period during which treatment with trypsin could rescue the cells on media containing low concentrations of K+. In an adenosinetriphosphate (ATP phohsphohydrolase, EC 3.6.1.3) deficient (uncA) mutant, survival after colicin treatment was observed at lower Mg2+ concentrations than those required by the wild type, and Rb+ could replace K+. Cells treated with colicin E1 (but not with colicin I2, E3, or Ib) also survived under conditions permitting survival of colicin K.

Mutants in three genes affecting transport of magnesium in Escherichia coli: genetics and physiology.

Mutants in three genes affecting two Mg2+ transport systems are described. System I, for which Co2+, Mn2+, and Mg2+ are substrates, is inactive in corA mutants corB mutants express system I after growth on high (10 mM) Mg2+ but not low (0.1 mM) Mg2+. Both corA and corB mutants are resistant to Co2+ or Mn2+. corA mutants are sensitive to CA2+. Transport system II is specific for Mg2+ and is repressed by growth on 10 mM Mg2+. mgt mutations inactivate system II. Growth on mgt mutants in normal except on very low (1 muM) concentrations of Mg2+, corA mgt strains exhibit no high-affinity, energy-dependent transport of Mg2+ and require 10 mM Mg2+ for optimal growth. The three genes are not linked. The corA locus is contransducible with ilv at 75 min, corB is cotransducible with pyrB at 85 min, and mgt is cotransducible with malB and mel at 81 min on the genetic map.

Cavernous hemangioma of the optic disc.

Cavernous hemangioma of the optic disc and retina is a vascular hamartoma composed of clusters of thin-walled saccular aneurysms partially filled with dark venous blood. It is now considered a neuro-oculo-cutaneous syndrome whose mode of inheritance is probably autosomal dominant. This report describes a cavernous hemangioma of the optic disc in a patient without skin or neurological findings. Its clinical appearance, generally benign ocular nature, and occasional association with central nervous system or cutaneous lesions is emphasized.