A Ca2+/cam protein kinase associated with Ca2+ transport in sarco(endo)plasmic vesicles from tracheal smooth muscle

Immunohistochemical Localization of Ca2+/Calmodulin-dependent Protein Kinase IV in Outer Hair Cells

“…Of these, CaM kinase IV has been implicated in the regulation of gene transcription based, in part, on its localization in nuclei of cerebellar granular cells (Ohmstede et al 1989;Jensen et al 1991;Sakagami et al 1994;Nakamura et al 1995) and in neurotransmitter release based on its capacity to phosphorylate synapsin I (Miyano et al 1992). In addition, CaM kinases have been found to act by phosphorylating and thereby increasing the activity of SERCAs in sarcoplasmic reticulum (Leddy et al 1993;Mattiazzi et al 1994;Baltas et al 1995;Damiani et al 1995;de Alfonzo et al 1996;Grover et al 1996;Narayanan and Xu 1997).…”

Two Guanylylcyclases Regulate the Muscarinic Activation of Airway Smooth Muscle

“…This 60-s cGMP signal is a product of NPR-GC-B as above mentioned in references [35,54,55]. Recently, we further recognized that muscarinic agonist and mastoparan activations of NPR-GC-B in isolated BTSM plasma membranes fraction involve this mastoparan-sensitiveGq16 protein in [54]. Thus, the pre-exposure of TSM strips to mastoparan stimulated the Gq16 protein, disrupting this signal cascade inducing the failure of this muscarinic-dependent TSM contraction.…”

“…Recently, it has been suggested that a plasma membrane-bound GC, may provide a localized pool of cGMP in [68], which seems to be in a similar trend suggested by our work. The second signal peak is a product of a the activation of M3AChR coupled to the stimulation of mastoparan-sensitive Gq16 as reported in reference [54] to turn on a transmembrane-homodimer as NPR-GC-B as described in [35,55].…”

“…Identification of 1/ 1 sGC heterodimer in membrane fractions isolated from BTSM strips under muscarinic agonist exposure. Isolated BTSM were incubated in KRB at 37°C in the presence and absence of muscarinic agonist CC (1 x10 −5 M) during 0, 20 s and 60 s. The samples were immediately frozen and pulverized with a mortar in liquid nitrogen, following the protocols described to isolate crude plasma membranes fraction as described in [31,54]. The final membranes sediments were electrophoresed in 12% PAGE-SDS, transferred to nitrocellulose membranes, and probed with specific antibodies against the 1, 2, (A) 1 and 2 (B) of NO-sGC subunits.…”

“…After each 10 s, the BTSM strips were removed and immediately frozen in liquid N2 and processed to prepare crude membranes fraction as described in [54]. In these membrane sediments, GC assays were performed in duplicate as described previously in [53].…”