Advanced

Bradykinin and bombesin rapidly stimulate tyrosine phosphorylation of a 120-kDa group of proteins in Swiss 3T3 cells

Leeb-Lundberg, L. M Fredrik LU and Song, Xin Hua (1991) In Journal of Biological Chemistry 266(12). p.7746-7749
Abstract

We have examined the effect of bradykinin (BK) and other peptide mediators with related cellular actions on tyrosine phosphorylation in confluent Swiss 3T3 fibroblast cells using an anti-phosphotyrosine antibody. Immunoblots of extracts from cells stimulated with BK showed a major heterogeneous band centered at Mr 120,000. Three phosphorylated protein species were present within this band. The lower of these three phosphoproteins was occasionally present under basal conditions. The detection of this group of phosphoproteins by the antibody was prevented by coincubation with an excess of phosphotyrosine but not with an excess of phosphoserine or phosphothreonine. The BK-promoted increase in phosphorylation was rapid and... (More)

We have examined the effect of bradykinin (BK) and other peptide mediators with related cellular actions on tyrosine phosphorylation in confluent Swiss 3T3 fibroblast cells using an anti-phosphotyrosine antibody. Immunoblots of extracts from cells stimulated with BK showed a major heterogeneous band centered at Mr 120,000. Three phosphorylated protein species were present within this band. The lower of these three phosphoproteins was occasionally present under basal conditions. The detection of this group of phosphoproteins by the antibody was prevented by coincubation with an excess of phosphotyrosine but not with an excess of phosphoserine or phosphothreonine. The BK-promoted increase in phosphorylation was rapid and transient with the peak response apparent following BK exposure for 1 min. The response was dose-dependent with half-maximal effect occurring at 10-30 nM BK. The antagonist Arg0, Hyp3, Thi5,8, D-Phe7-BK completely inhibited the response indicating that BK was acting via a B2 kinin receptor. Bombesin, at 0.1 μM, stimulated an increase in phosphorylation of the 120-kDa group of proteins with the same efficacy as 0.1 μM BK. On the other hand, 1 μM vasopressin was considerably less efficaceous than either of the former agonists. Short-term preexposure to 0.1 μM 12-O-tetradecanoyl-phorbol-13-acetate (1 min), a protein kinase C stimulator, or 30 μM H7 (15 min), a protein kinase C inhibitor, had no significant effect either on the basal or BK-promoted increase in tyrosine phosphorylation of these proteins. BK also stimulated inositol phosphate formation in these cells. Genistein, a tyrosine kinase inhibitor, inhibited BK stimulation of tyrosine phosphorylation. In addition, genistein partially inhibited BK stimulation of inositol phosphate formation. These results show that an increase in tyrosine phosphorylation of a 120-kDa group of proteins is an early protein kinase C-independent cellular signal elicited by both bradykinin and bombesin.

(Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
266
issue
12
pages
7746 - 7749
publisher
ASBMB
external identifiers
  • pmid:2019598
  • scopus:0025744009
ISSN
0021-9258
language
English
LU publication?
no
id
c9bd9398-9dcc-4133-be11-daa543c2fc3b
alternative location
http://www.jbc.org/content/266/12/7746.abstract
date added to LUP
2019-06-12 11:49:18
date last changed
2019-11-25 09:31:23
@article{c9bd9398-9dcc-4133-be11-daa543c2fc3b,
  abstract     = {<p>We have examined the effect of bradykinin (BK) and other peptide mediators with related cellular actions on tyrosine phosphorylation in confluent Swiss 3T3 fibroblast cells using an anti-phosphotyrosine antibody. Immunoblots of extracts from cells stimulated with BK showed a major heterogeneous band centered at M<sub>r</sub> 120,000. Three phosphorylated protein species were present within this band. The lower of these three phosphoproteins was occasionally present under basal conditions. The detection of this group of phosphoproteins by the antibody was prevented by coincubation with an excess of phosphotyrosine but not with an excess of phosphoserine or phosphothreonine. The BK-promoted increase in phosphorylation was rapid and transient with the peak response apparent following BK exposure for 1 min. The response was dose-dependent with half-maximal effect occurring at 10-30 nM BK. The antagonist Arg<sup>0</sup>, Hyp<sup>3</sup>, Thi<sup>5,8</sup>, D-Phe<sup>7</sup>-BK completely inhibited the response indicating that BK was acting via a B2 kinin receptor. Bombesin, at 0.1 μM, stimulated an increase in phosphorylation of the 120-kDa group of proteins with the same efficacy as 0.1 μM BK. On the other hand, 1 μM vasopressin was considerably less efficaceous than either of the former agonists. Short-term preexposure to 0.1 μM 12-O-tetradecanoyl-phorbol-13-acetate (1 min), a protein kinase C stimulator, or 30 μM H7 (15 min), a protein kinase C inhibitor, had no significant effect either on the basal or BK-promoted increase in tyrosine phosphorylation of these proteins. BK also stimulated inositol phosphate formation in these cells. Genistein, a tyrosine kinase inhibitor, inhibited BK stimulation of tyrosine phosphorylation. In addition, genistein partially inhibited BK stimulation of inositol phosphate formation. These results show that an increase in tyrosine phosphorylation of a 120-kDa group of proteins is an early protein kinase C-independent cellular signal elicited by both bradykinin and bombesin.</p>},
  author       = {Leeb-Lundberg, L. M Fredrik and Song, Xin Hua},
  issn         = {0021-9258},
  language     = {eng},
  month        = {04},
  number       = {12},
  pages        = {7746--7749},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Bradykinin and bombesin rapidly stimulate tyrosine phosphorylation of a 120-kDa group of proteins in Swiss 3T3 cells},
  url          = {http://www.jbc.org/content/266/12/7746.abstract},
  volume       = {266},
  year         = {1991},
}