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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 115 (1990), S. 41-50 
    ISSN: 1432-1424
    Keywords: inward rectification ; voltage-dependent K+ current ; Ca2+-dependent K+ current ; Paramecium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Hyperpolarization of voltage-clampedParamecium tetraurelia in K+ solutions elicits a complex of Ca2+ and K+ currents. The tail current that accompanies a return to holding potential (−40 mV) contains two K+ components. The tail current elicited by a step to −110 mV of ≥50-msec duration contains fast-decaying (τ≈3.5 msec) and slow-decaying (τ≈20 msec) components. The reversal potential of both components shifts by 55–57 mV/10-fold change in external [K+], suggesting that they represent pure K+ currents. The dependence of the relative amplitudes of the two tail currents on duration of hyperpolarization suggests that the slow K+ current activates slowly and is sustained, whereas the fast current activates rapidly during hyperpolarization and then rapidly inactivates. Iontophoretic injection of a Ca2+ chelator, EGTA, specifically reduces slow tail-current amplitude without affecting the fast tail component. Both K+ currents are inhibited by extracellular TEA+ in a concentration-dependent, noncooperative manner, whereas the fast K+ current alone is inhibited by 0.7mm quinidine.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1424
    Keywords: calmodulin ; mutation ; ion currents ; Paramecium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Two behavioral mutants ofParamecium tetraurelia, pantophobiacs A1 and A2, have single amino acid defects in the structure of calmodulin. The mutants exhibit several major ion current defects under voltage clamp: (i) the Ca2+-dependent K+ current activated upon depolarization ofParamecium is greatly reduced or missing in both mutants, (ii) both mutants lack a Ca2+-dependent K+ current activated upon hyperpolarization, and (iii) the Ca2+-dependent Na+ current is significantly smaller in pantophobiac A1 compared with the wild type, whereas this current is slightly increased in pantophobiac A2. Other, minor defects include a reduction in peak amplitude of the depolarization-activated Ca2+ current in pantophobiac A2, increased rates of voltage-dependent inactivation of this Ca2+ current in both pantophobiac A1 and pantophobiac A2, and an increase in the time required for the hyperpolarization-activated Ca2+ current to recover from inactivation in the pantophobiacs. The diversity of the pantophobiac mutations' effects on ion current function may indicate specific associations of calmodulin with a variety of Ca2+-related ion channel species inParamecium.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1424
    Keywords: calmodulin ; Ca2+-dependent K+ channels ; ion channel regulation ; mutations ; Paramecium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Paramecium tetraurelia possesses two Ca2+-dependent K+ currents, activated upon depolarizationI K(Ca,d), or upon hyperpolarizationI K(Ca,h). The two currents are mediated by pharmacologically distinct ion channel populations. Three mutations ofP. tetraurelia affect these current.s Pantophobiac A mutations (pntA) cause calmodulin sequence defects, resulting in the loss of both Ca2+-dependent K+ currents. A second mutation, TEA-insensitive A (teaA), greatly enhancesI K(Ca,d) but has no affect onI K(Ca,h). A third mutation,restless (rst), also increasesI K(Ca,d) slightly, but its principle effect is in causing an early activation ofI K(Ca,h). Interactions between the products of these three genes were investigated by constructing three double mutants. BothteaA andrst restoreI K(Ca,d) andI K(Ca,h) in pantophobiac A1, but the phenotypes ofteaA andrst are not corrected by a second mutation. These observations may indicate a role for the gene products ofteaA andrst in regulating the activity ofI K(Ca,d) andI K(Ca,h), respectively.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 12 (1992), S. 365-371 
    ISSN: 0887-3585
    Keywords: Amino acid sequence/methylated lysines/protein conformation/Ca+ +-dependent K+ channel/cam behavioral mutants ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Paramecium tetraurelia behavioral mutant cam12 displays a “fast-2” behavioral phenotype: it fails to respond to Na+ stimuli. Electrophysiologically, it lacks a Ca+ +-dependent Na+ current. Genetics and DNA sequencing showed the primary defect of cam12 to be in the calmodulin gene (Kink et al., 1990). To correlate calmodulin structure and function in Paramecium, we elucidated the primary structure of cam12 calmodulin. Peptide sequencing confirmed the two point mutations predicted by the DNA sequence: a glycine-to-glutamate substitution at position 40 and an aspartate-to-asparagine substitution at position 50. Our results further showed that lysine 13 and lysine 115 were methylated normally in cam12. It is likely that the electrophysiological abnormalities of cam12 are a direct reflection of the amino-acid substitutions, as opposed to improper posttranslational modification.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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