Hyper-Activation of pp60(Src) Limits Nitric Oxide Signaling by Increasing Asymmetric Dimethylarginine Levels During Acute Lung Injury

Authors

Sanjiv Kumar
Xutong Sun
Satish Kumar Noonepalle
Qing Lu
Evgeny Zemskov
Ting Wang
Saurabh Aggarwal
Christine Gross
Shruti Sharma
Ankit A. Sesai
John D. Catravas, Old Dominion University

Document Type

Article

Publication Date

1-2017

Publication Title

Free Radical Biology and Medicine

Volume

102

Pages

217-228

DOI

10.1016/j.freeradbiomed.2016.11.008

Abstract

The molecular mechanisms by which the endothelial barrier becomes compromised during lipopolysaccharide (LPS) mediated acute lung injury (ALI) are still unresolved. We have previously reported that the disruption of the endothelial barrier is due, at least in part, to the uncoupling of endothelial nitric oxide synthase (eNOS) and increased peroxynitrite-mediated nitration of RhoA. The purpose of this study was to elucidate the molecular mechanisms by which LPS induces eNOS uncoupling during ALI. Exposure of pulmonary endothelial cells (PAEC) to LPS increased pp60^Src activity and this correlated with an increase in nitric oxide (NO) production, but also an increase in NOS derived superoxide, peroxynitrite formation and 3-nitrotyrosine (3-NT) levels. These effects could be simulated by the over-expression of a constitutively active pp60^Src (Y527FSrc) mutant and attenuated by over-expression of dominant negative pp60^Src mutant or reducing pp60^Src expression. LPS induces both RhoA nitration and endothelial barrier disruption and these events were attenuated when pp60^Src expression was reduced. Endothelial NOS uncoupling correlated with an increase in the levels of asymmetric dimethylarginine (ADMA) in both LPS exposed and Y527FSrc over-expressing PAEC. The effects in PAEC were also recapitulated when we transiently over-expressed Y527FSrc in the mouse lung. Finally, we found that the pp60^Src-mediated decrease in DDAH activity was mediated by the phosphorylation of DDAH II at Y207 and that a Y207F mutant DDAH II was resistant to pp60^Src-mediated inhibition. We conclude that pp60^Src can directly inhibit DDAH II and this is involved in the increased ADMA levels that enhance eNOS uncoupling during the development of ALI.

Comments

NOTE: This is the author's manuscript version (peer-reviewed and accepted for publication) of a work that was published in Free Radical Biology and Medicine.The final version was published in final edited form as:

Kumar, S., Sun, X., Noonepalle, S. K., Lu, Q., Zemskov, E., Wang, T., … Black, S. M. (2017). Hyper-activation of pp60^Src limits nitric oxide signaling by increasing asymmetric dimethylarginine levels during acute lung injury. Free Radical Biology & Medicine, 102, 217–228. http://doi.org/10.1016/j.freeradbiomed.2016.11.008

Available at: http://doi.org/10.1016/j.freeradbiomed.2016.11.008

Original Publication Citation

Kumar, S., Sun, X. T., Noonepalle, S. K., Lu, Q., Zemskov, E., Wang, T., . . . Black, S. M. (2017). Hyper-activation of pp60^src limits nitric oxide signaling by increasing asymmetric dimethylarginine levels during acute lung injury. Free Radical Biology & Medicine, 102, 217-228. doi:10.1016/j.freeradbiomed.2016.11.008

Repository Citation

Kumar, Sanjiv; Sun, Xutong; Noonepalle, Satish Kumar; Lu, Qing; Zemskov, Evgeny; Wang, Ting; Aggarwal, Saurabh; Gross, Christine; Sharma, Shruti; Sesai, Ankit A.; and Catravas, John D., "Hyper-Activation of pp60(Src) Limits Nitric Oxide Signaling by Increasing Asymmetric Dimethylarginine Levels During Acute Lung Injury" (2017). Bioelectrics Publications. 153.
https://digitalcommons.odu.edu/bioelectrics_pubs/153