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N M Greene

Publications and source records attributed to N M Greene.

At least 19 recordsLinked to original sources

Identification of protein kinase C phosphorylation sites on bovine rhodopsin.

The protein kinase C phosphorylation sites on bovine rhodopsin were identified using proteolytic, phosphoamino acid, mass spectrometric, and peptide sequencing analyses. Tryptic removal of the 9 carboxyl-terminal residues of rhodopsin revealed that a major fraction of the phosphates incorporated by protein kinase C are in a region containing Ser334, Thr335, and Thr336. Phosphoamino acid analysis of the tryptic product established that Ser334 accounts for approximately 65% of the phosphorylation in this region. Analysis of the endoproteinase Asp-N-generated carboxyl terminus of rhodopsin by mass spectrometry and peptide sequencing revealed that Ser338 is also a primary phosphorylation site, with minor phosphorylation of Ser343. Quantitation of high pressure liquid chromatography-separated phosphopeptides, taken together with phosphoamino acid analysis of the tryptic product, revealed that Ser334 and Ser338 were phosphorylated equally and each accounted for approximately 35% of the total phosphorylation; Thr335/336 accounted for just under 20% of the phosphorylation, and Ser343 accounted for 10%. Thus, the primary protein kinase C sites are Ser334 and Ser338, with minor phosphorylation of Thr335/336 and Ser343. Ser334 and Ser338 have recently been identified as the primary sites of phosphorylation of rhodopsin in vivo (Ohguro, H., Van Hooser, J. P., Milam, A. H., and Palczewski, K. (1995) J. Biol. Chem. 270, 14259-14262). Of these sites, only Ser338 is a significant substrate for rhodopsin kinase in vitro. Identification of Ser334 as a primary protein kinase C target in vitro is consistent with protein kinase C modulating the phosphorylation of this site in vivo.

Amino Acid Sequence↗

Kinetics and localization of the phosphorylation of rhodopsin by protein kinase C.

Protein kinase C isolated from retina catalyzes the stoichiometric phosphorylation of bovine rhodopsin. Enzymological studies using receptor in rod outer segment membranes stripped of peripheral proteins reveal that the phosphorylation is independent of receptor conformation or liganded state; the half-time for phosphorylation of unbleached (dark-adapted) rhodopsin, bleached (light-activated) rhodopsin, and opsin (chromophore removed) is the same. The phosphorylation by protein kinase C is Ca2+ and lipid regulated; the Km for Ca2+ decreases with increasing concentrations of membrane, consistent with known properties of Ca(2+)-regulated protein kinase Cs. The Km for ATP is 27 microM, with an optimal concentration for MgCl2 of approximately 1 mM. The phosphorylation of rhodopsin by protein kinase C is inhibited by the protein kinase C-selective inhibitor sangivamycin. Proteolysis by Asp-N reveals that all the protein kinase C phosphorylation sites are on the carboxyl terminus of the receptor. Cleavage with trypsin indicates that Ser338, the primary phosphorylation site of rhodopsin kinase, is not phosphorylated significantly; rather, the primary phosphorylation site of protein kinase C is on the membrane proximal half of the carboxyl terminus. The protein kinase C-catalyzed phosphorylation of rhodopsin is analogous to the ligand-independent phosphorylation of other G protein-coupled receptors that is catalyzed by second messenger-regulated kinases.

Amino Acid Sequence↗

Here we go again.

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Anesthetics↗

The 31st Rovenstine Lecture. The changing horizons in anesthesiology.

Modern anesthesiology differs widely from what it was 40-50 years ago, not only because of what anesthesiology now involves in the operating room, but also because anesthesiology has expanded its horizons and activities above and beyond the provision of surgical anesthesia. These changes and the identity of modern anesthesiology are, however, but poorly understood, if understood at all, by the majority of laity and physicians alike. Such lack of identity, especially in the minds of those at the policy- and decision-making level, can only endanger the vitality and future of anesthesiology in an era of sweeping changes in health care-delivery systems. The problem of public identity of our specialty includes the historically correct, but, contemporaneously, all too often misleading name of our specialty. It is suggested that it is appropriate, at this time, to at least consider the potential advantages of changing the name of our specialty to, say, metesthesiology and metesthesiologist, to indicate that while, today, our specialty continues to involve operative anesthesia, it extends above and beyond to include a wide variety of professional activities outside the operating room richly rewarding to patient and practitioner alike.

Anesthesiology↗

An American anesthesia training program in East Africa.

A unique 5-year program designed to improve the quality and availability of anesthesia care in developing nations has been initiated in East Africa. Based upon North American anesthesiologists serving as volunteer teachers, the program emphasizes teaching to the exclusion of taking over anesthesia care. The teaching is practical and relevant to the clinical practice of anesthesia in developing countries. The volunteers teachers serve, one at a time, 12 months of the year for several years in two pre-existing structured East African anesthesia training programs, thereby providing the advantages of consistent, reliable teaching assistance for a period of years to programs of demonstrable interest and experience in anesthesia training.

Africa, Eastern↗

Zones of differential sensory block during extradural anaesthesia.

We have measured spinal segmental levels of anaesthesia to light touch (LT), pinprick (PP) and cold temperature discrimination (TE) during 2% lignocaine extradural anaesthesia in 22 patients, to determine if zones of differential sensory block develop during extradural anaesthesia and, if so, the extent to which TE extends beyond PP or LT levels and how age affects differential block. The median thoracic dermatomal levels were 4.5 for LT, 2.0 for PP and 2.0 for TE. Zones of differential sensory block developed within 5 min of extradural injection of local anaesthetic, and persisted for the next 55 min. In all instances, PP extended more cephalad than LT, and TE extended above PP levels. There were no differences in the extent of zones between the two groups of patients with mean ages of 28 and 48 yr. Thus, during extradural anaesthesia, sympathetic denervation extended one to two spinal segments above the sensory levels of LT and PP anaesthesia, age (28 vs 48 yr) affected neither the cephalad extent nor the width of zones of differential block, and PP levels of anaesthesia were closer to presumed levels of sympathetic block than were LT levels.

Adult↗

Time-courses of zones of differential sensory blockade during spinal anesthesia with hyperbaric tetracaine or bupivacaine.

The purposes of this study were twofold: to compare bupivacaine and tetracaine spinal anesthesia with regard to the zones of differential sensory blockade and to evaluate the time-courses of the widths of the zones of differential sensory blockade during spinal anesthesia. In 51 patients, the most rostral levels of sensory denervation to light touch, pinprick, and temperature discrimination were measured. There was no statistically significant difference in the height of sensory blockade in the 29 patients given bupivacaine and in the 22 patients given equipotent doses of tetracaine. The widths of the zones of differential blockade were also not statistically different between the two groups during onset, maintenance, and regression of anesthesia, except that the light touch-to-pinprick and light touch-to-temperature zones of differential blockade were greater with bupivacaine than with tetracaine 30 min after subarachnoid injection. The width of the zones of differential blockade also remained unchanged within each group during onset, maintenance, and regression of anesthesia. Changes in, and absolute levels of, blood pressure and heart rate were similar with both bupivacaine and tetracaine throughout. We conclude that zones of differential sensory blockade are essentially the same with tetracaine and bupivacaine, that the widths of the zones of differential sensory blockade remain constant during onset, maintenance, and offset of spinal anesthesia, and that bupivacaine and tetracaine are associated with similar changes in heart rate and blood pressure during spinal anesthesia.

Adult↗