PMID- 10514519
OWN - NLM
STAT- MEDLINE
DCOM- 19991119
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 42
DP  - 1999 Oct 15
TI  - Neuronal Ca(2+) sensor 1. Characterization of the myristoylated protein, its
      cellular effects in permeabilized adrenal chromaffin cells, Ca(2+)-independent
      membrane association, and interaction with binding proteins, suggesting a role in
      rapid Ca(2+) signal transduction.
PG  - 30258-65
AB  - Overexpression of frequenin and its orthologue neuronal Ca(2+) sensor 1 (NCS-1)
      has been shown to increase evoked exocytosis in neurons and neuroendocrine cells.
      The site of action of NCS-1 and its biochemical targets that affect exocytosis
      are unknown. To allow further investigation of NCS-1 function, we have
      demonstrated that NCS-1 is a substrate for N-myristoyltransferase and generated
      recombinant myristoylated NCS-1. The bacterially expressed NCS-1 shows
      Ca(2+)-induced conformational changes. The possibility that NCS-1 directly
      interacts with the exocytotic machinery to enhance exocytosis was tested using
      digitonin-permeabilized chromaffin cells. Exogenous NCS-1 was retained in
      permeabilized cells but had no effect on Ca(2+)-dependent release of
      catecholamine. In addition, exogenous NCS-1 did not regulate cyclic nucleotide
      levels in this system. These data suggest that the effects of NCS-1 seen in
      intact cells are likely to be due to an action on the early steps of
      stimulus-secretion coupling or on Ca(2+) homeostasis. Myristoylated NCS-1 bound
      to membranes in the absence of Ca(2+) and endogenous NCS-1 was tightly
      membrane-associated. Using biotinylated NCS-1, a series of specific binding
      proteins were detected in cytosol, chromaffin granule membrane, and microsome
      fractions of adrenal medulla. These included proteins distinct from those
      detected by biotinylated calmodulin, demonstrating the presence of multiple
      specific Ca(2+)-independent and Ca(2+)-dependent binding proteins as putative
      targets for NCS-1 action. A model for NCS-1 function, from these data, indicates 
      a constitutive membrane association independent of Ca(2+). This differs from the 
      Ca(2+) myristoyl switch model for the closely related recoverin and suggests a
      possible action in rapid Ca(2+) signal transduction in response to local Ca(2+)
      signals.
FAU - McFerran, B W
AU  - McFerran BW
AD  - The Physiological Laboratory, University of Liverpool, Crown Street, Liverpool
      L69 3BX, United Kingdom.
FAU - Weiss, J L
AU  - Weiss JL
FAU - Burgoyne, R D
AU  - Burgoyne RD
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Calcium-Binding Proteins)
RN  - 0 (DNA Primers)
RN  - 0 (Neuronal Calcium-Sensor Proteins)
RN  - 0 (Neuropeptides)
RN  - 0 (Recombinant Proteins)
RN  - 0 (frequenin calcium sensor proteins)
SB  - IM
MH  - Adrenal Medulla/cytology/*metabolism
MH  - Animals
MH  - Base Sequence
MH  - Brain/metabolism
MH  - *Calcium Signaling
MH  - Calcium-Binding Proteins/*metabolism
MH  - Cattle
MH  - Chromaffin Cells/*metabolism
MH  - DNA Primers
MH  - Neuronal Calcium-Sensor Proteins
MH  - Neuropeptides/*metabolism
MH  - Protein Binding
MH  - Rats
MH  - Rats, Wistar
MH  - Recombinant Proteins/metabolism
EDAT- 1999/10/09 00:00
MHDA- 1999/10/09 00:01
CRDT- 1999/10/09 00:00
PHST- 1999/10/09 00:00 [pubmed]
PHST- 1999/10/09 00:01 [medline]
PHST- 1999/10/09 00:00 [entrez]
AID - 10.1074/jbc.274.42.30258 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Oct 15;274(42):30258-65. doi: 10.1074/jbc.274.42.30258.