Control of Vascular Permeability and Vascular Smooth Muscle in the Respiratory Tract by Multiple Neuropeptides

I In n v vi iv vo o e ex xp po os su ur re e t to o n ni it tr ro og ge en n d di io ox xi id de e ( (N NOO2 2 ) ) i in nd du uc ce es s a a d de ec cr re ea as se e i in n c ca al lc ci it to on ni in n g ge en ne e--r re el la at te ed d p pe ep pt ti id de e ( (C CG GR RP P) ) a an nd d t ta ac ch hy yk ki in ni in n i im mm mu un no or re ea ac c--t ti iv vi it ty y i in n g gu ui in ne ea a--p pi ig g p pe er ri ip ph he er ra al l a ai ir rw wa ay ys s We examined the effects of inhaled nitrogen dioxide (NO 2 ), a well-known indoor and outdoor air pollutant, on pulmonary sensory neuropeptides. Guinea-pigs were exposed for 4 h to 18 parts per million (ppm) NO 2 or to air (n=5 each). At the end of the exposure, they were killed with urethane and their lungs were fixed in 1% paraformaldehyde in phosphate-buffered saline. Cryostat sections were stained with antisera to an anatomical nerve marker, protein gene product (PGP) 9.5, and to CGRP and tachykinins, utilizing the avidin-biotinylated peroxidase method.In the noncartilaginous airways (diameter <250 µm) of NO 2 -exposed animals, less tachykinin-and CGRP-immunoreactive nerve fibres were found compared with controls. No change was seen in the total nerve fibre distribution (PGP 9.5).It is concluded that the peptidergic nerves of guinea-pig peripheral airways are a sensitive indicator of exposure to nitrogen dioxide.

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“…This finding is not surprising since SP, other tachykinins and CGRP are known to be co-localized in some sensory nerves [32,33].…”

Section: Discussionmentioning

confidence: 70%

In vivo exposure to nitrogen dioxide (NO2) induces a decrease in calcitonin gene-related peptide (CGRP) and tachykinin immunoreactivity in guinea-pig peripheral airways

Lucchini

Springall²,

Chitano³

et al. 1996

Eur Respir J

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“…The undecapeptide SP belongs to the tachykinin neurotransmitter family. [23,24] It is distributed both in the peripheral nervous system, [25] where it mediates different physiological responses, including smooth muscle contraction, [26] vasodilation, [27] and increase in vascular permeability, [28] as well as in the central nervous system, [29] where it is involved mainly in inflammatory response [30] and pain perception. [31] SP action is regulated through its binding to the specific neurokinin-1 receptor (NK-1R), a G-proteincoupled transmembrane protein.…”

Section: Introductionmentioning

confidence: 99%

Deamidation and Transamidation of Substance P by Tissue Transglutaminase Revealed by Electron‐Capture Dissociation Fourier Transform Mass Spectrometry

Fornelli

Schmid

Grasso

et al. 2010

Chemistry A European J

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Tissue transglutaminase (tTGase) catalyzes both deamidation and transamidation of peptides and proteins by using a peptidyl glutamine as primary substrate. A precise consensus sequence for the enzyme is unknown and the ratio between deamidated and transamidated (or cross-linked) reaction products is highly substrate-dependent. Due to its overlapping body distribution with tTGase and ease of manipulation with tandem mass spectrometry, we used the neuropeptide substance P as a model to investigate the associated enzymatic kinetics and reaction products. Online liquid-chromatography Fourier-transform ion-cyclotron-resonance mass spectrometry (FT-ICR MS) combined with electron-capture dissociation (ECD) was employed to study the tTGase-induced modifications of substance P. A particular strength of ECD for peptide-enzyme reaction product monitoring is its ability to distinguish isomeric amino acids, for example, Glu and iso-Glu, by signature product ions. Our studies show that the primary reaction observed is deamidation, with the two consecutive glutamine residues converted sequentially into glutamate: first Gln(5) , and subsequently Gln(6) . We then applied ECD FT-ICR MS to identify the transamidation site on an enzymatically cross-linked peptide, which turned out to correspond to Gln(5) . Three populations of substance-P dimers were detected that differed by the number of deamidated Gln residues. The higher reactivity of Gln(5) over Gln(6) was further confirmed by cross-linking SP with monodansylcadaverine (MDC). Overall, our approach described herein is of a general importance for mapping both enzymatically induced post-translational protein modifications and cross-linking. Finally, in vitro Ca-signaling assays revealed that the main tTGase reaction product, the singly deamidated SP (RPKPEQFFGLM-NH(2) ), has increased agonist potency towards its natural receptor, thus confirming the biologically relevant role of deamidation.

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Control of Vascular Permeability and Vascular Smooth Muscle in the Respiratory Tract by Multiple Neuropeptides

Cited by 2 publications

References 18 publications

In vivo exposure to nitrogen dioxide (NO2) induces a decrease in calcitonin gene-related peptide (CGRP) and tachykinin immunoreactivity in guinea-pig peripheral airways

In vivo exposure to nitrogen dioxide (NO2) induces a decrease in calcitonin gene-related peptide (CGRP) and tachykinin immunoreactivity in guinea-pig peripheral airways

Deamidation and Transamidation of Substance P by Tissue Transglutaminase Revealed by Electron‐Capture Dissociation Fourier Transform Mass Spectrometry

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