N-terminal myristoylation is required for membrane localization of cGMP-dependent protein kinase type II

The apical membrane of intestinal epithelial cells harbors a unique isozyme of cGMP-dependent protein kinase (cGK type II) which acts as a key regulator of ion transport systems, including the cystic fibrosis transmembrane conductance regulator (CFTR)-chloride channel. To explore the mechanism of cGK II membrane-anchoring, recombinant cGK II was expressed stably in HEK 293 cells or transiently in COS-1 cells. In both cell lines, cGK II was found predominantly in the particulate fraction. Immunoprecipitation of solubilized cGK II did not reveal any other tightly associated proteins, suggesting a membrane binding motif within cGK II itself. The primary structure of cGK II is devoid of hydrophobic transmembrane domains; cGK II does, however, contain a penultimate glycine, a potential acceptor for a myristoyl moiety. Metabolic labeling showed that cGK II was indeed able to incorporate [3H]myristate. Moreover, incubation of cGK II-expressing 293 cells with the myristoylation inhibitor 2-hydroxymyristic acid (1 mM) significantly increased the proportion of cGK II in the cytosol from 10 +/- 5 to 35 +/- 4%. Furthermore, a nonmyristoylated cGK II Gly2 --> Ala mutant was localized predominantly in the cytosol after transient expression in COS-1 cells. The absence of the myristoyl group did not affect the specific enzyme activity or the Ka for cGMP and only slightly enhanced the thermal stability of cGK II. These results indicate that N-terminal myristoylation fulfills a crucial role in directing cGK II to the membrane

EXTRACTION OF LINES AND REGIONS FROM GREY TONE LINE DRAWING IMAGES

An algorithm is described for extracting lines from grey level digitizations of industrial drawings. The algorithm is robust, non iterati e, and sequential, and includes procedures for differentiating shaded areas from lines. Examples are given for complex regions of a typical mechanical drawing

Feminist phenomenology and the woman in the running body

Modern phenomenology, with its roots in Husserlian philosophy, has been taken up and utilised in a myriad of ways within different disciplines, but until recently has remained relatively under-used within sports studies. A corpus of sociological-phenomenological work is now beginning to develop in this domain, alongside a longer standing literature in feminist phenomenology. These specific social-phenomenological forms explore the situatedness of lived-body experience within a particular social structure. After providing a brief overview of key strands of phenomenology, this article considers some of the ways in which sociological, and particularly feminist phenomenology, might be used to analyse female sporting embodiment. For illustrative purposes, data from an autophenomenographic project on female distance running are also included, in order briefly to demonstrate the application of phenomenology within sociology, as both theoretical framework and methodological approach

Subjective wellbeing among young dancers with disabilities

Little is known about the subjective wellbeing (SWB) of young dancers with disabilities and whether it changes over time. The aim of this study was to assess the SWB of young dancers with disabilities enrolled on an extracurricular inclusive talent development programme in the UK at two time points. Twenty-two young dancers completed the Personal Wellbeing Index for people with intellectual disability at the beginning of the academic year. Thirteen dancers completed the questionnaire a second time towards the end of the academic year. Scores were compared with normative values, and a Wilcoxon Signed Rank test was conducted to assess change over time. The participants reported high levels of SWB at both time points in comparison with normative values. There was no significant change in wellbeing scores over time. The study contributes to a growing body of literature suggesting that people with disabilities have high levels of SWB. Although causality cannot be assumed, inclusive dance programmes may contribute to SWB and allow young people with disabilities to overcome the barriers associated with physical activity