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Ernesto Carafoli

Publications and source records attributed to Ernesto Carafoli.

25 records · Page 2Linked to original sources
PubMed2002

Calcium signaling: a tale for all seasons.

An experiment performed in London nearly 120 years ago, which by today's standards would be considered unacceptably sloppy, marked the beginning of the calcium (Ca(2+)) signaling saga. Sidney Ringer [Ringer, S. (1883) J. Physiol. 4, 29-43] was studying the contraction of isolated rat hearts. In earlier experiments, Ringer had suspended them in a saline medium for which he admitted to having used London tap water, which is hard: The hearts contracted beautifully. When he proceeded to replace the tap water with distilled water, he made a startling finding: The beating of the hearts became progressively weaker, and stopped altogether after about 20 min. To maintain contraction, he found it necessary to add Ca(2+) salts to the suspension medium. Thus, Ringer had serendipitously discovered that Ca(2+), hitherto exclusively considered as a structural element, was active in a tissue that has nothing to do with bone or teeth, and performed there a completely novel function: It carried the signal that initiated heart contraction. It was a landmark observation, which should have immediately aroused wide interest. Unexpectedly, however, for decades it attracted no particular attention. Occasionally, farsighted pioneers argued forcefully for a messenger role of Ca(2+), offering compelling experimental evidence. Among them, one could quote L. V. Heilbrunn [Heilbrunn, L. V. (1940) Physiol. Zool. 13, 88-94], who contracted frog muscle fibers by applying Ca(2+) salts to their cut ends, but not to their surfaces. Heilbrunn correctly concluded that Ca(2+) had diffused from the cut ends to the internal contractile elements to elicit their contraction. One could also quote K. Bailey [Bailey, K. (1942) Biochem. J. 36, 121-139], who showed that the ATPase activity of myosin was strongly activated by Ca(2+) (but not by Mg(2+)), and concluded that the liberation of Ca(2+) in the neighborhood of the myosin controlled muscle contraction. Clearly, enough evidence was there, but only a handful of people had the vision to see it and to foresee its far-reaching implications. Perhaps no better example of clairvoyance can be offered than the quip by O. Loewy in 1959: "Ja Kalzium, das ist alles!"

Animals↗
PubMed2002

Differential membrane targeting of the SERCA and PMCA calcium pumps: experiments with recombinant chimeras.

Structural features underlying retention of the SERCA pump in intracellular compartments and the sorting of the PMCA pump to the plasma membrane are not known. The biochemical properties of the two pumps suggest that their differential localization may respond to specific functional demands. The two pumps may control Ca(2+) gradients of different magnitude and dynamic properties. For instance, it has recently become clear that the Ca(2+) gradient across the endoplasmic reticulum (ER) membrane is smaller than that across the plasma membrane. Previous experiments with chimerical constructs of the SERCA and PMCA pumps had suggested a role for the amino-terminal domain in the ER retention of the SERCA pump. Experiments aimed at narrowing down the region responsible for the retention now indicate that the first 28 amino acids of the SERCA pump may play a role in membrane localization. Results also suggest that the formation of oligomers (possibly through the first 28 amino acids) might be critical to the retention mechanism.

Adenosine Triphosphate↗
PubMed2002

A study of the activity of the plasma membrane Na/Ca exchanger in the cellular environment.

The Na/Ca exchanger is a large-capacity system that is supposed to be predominant in the ejection of calcium in excitable tissues. Its activity is normally evaluated in electro-physiological experiments or using Na-loaded cells or plasma membrane vesicles in experiments in which the activity is estimated by measuring the uptake of isotopic calcium. Here the activity of the exchanger has been evaluated in model cells that coexpress the exchanger together with the (protein) Ca indicator aequorin targeted to the different subcellular compartments. Exchanger isoform 1 (NCX1) has been used for experiments in which the effect of its overexpression has been evaluated on the Ca homeostasis in the cytoplasm at large, in the endoplasmic reticulum, in mitochondria, and in the layer of cytoplasm underneath the plasma membrane. The experiments have shown that, despite its very low calcium affinity, the exchanger was active in cells at rest in which calcium is around 100 nM. They have also shown that expressed NCX1 significantly reduced the cytosolic and mitochondrial calcium transients produced by the emptying of the ER calcium stores with InsP3-linked agonists. The effect of the expressed NCX1 on the calcium homeostasis in the domain underneath the plasma membrane was more complex: The exchanger apparently catalyzed a reverse operation, leading to increased penetration of calcium.

Adenosine Triphosphate↗
PubMedDate not supplied

Exporting calcium from cells.

All eukaryotic cells import Ca2+ through a number of variously gated plasma membrane channels. Once inside cells, Ca2+ transmits information to a large number of (enzyme) targets. Eventually, it must be exported again, to prevent the overloading of the cytosol with Ca2+. Two systems export Ca2+ from cells: a high affinity, low capacity Ca2+-ATPase, and a lower affinity, but much larger capacity, Na+/Ca2+ exchanger. The ATPase (commonly called the Ca2+ pump) is the fine-tuner of cell Ca2+, as it functions well even if the concentration of the ion drops below the microM level. It is a large enzyme, with 10 transmembrane domains and a C-terminal cytosolic tail that contains regulatory sites, including a calmodulin-binding domain. Four distinct gene products plus a large number of splice variants have been described. Some are tissue specific, the isoform 2 being specifically expressed in the sensorial cells of the Corti organ in the inner-ear. Its genetic absence causes deafness in mice. Two different families of the Na+/Ca2+ exchanger exist, one of which, originally described in photoreceptors, transports K+ and Ca2+ in exchange for Na+. The exchanger is particularly active in excitable cells, e.g., heart, where the necessity cyclically arises to rapidly eject large amounts of Ca2+. In addition to heart, the exchanger is particularly important to neurons: the cleavage of the most important neuronal isoform (NCX3) by calpains activated by excitotoxic treatments generates Ca2+ overload and eventually cell death.

Animals↗