Detection of NADPH-diaphorase activity in Paramecium primaurelia

Andrea Amaroli, Marzia Ognibene, Francesca Trielli, Sonya Trombino, Carla Falugi, Maria Umberta Delmonte Corrado

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Recently, we showed that Paramecium primaurelia synthesizes molecules functionally related to the cholinergic system and involved in modulating cell-cell interactions leading to the sexual process of conjugation. It is known that nitric oxide (NO) plays a role in regulating the release of transmitter molecules, such as acetylcholine, and that the NO biosynthetic enzyme, nitric oxide synthase (NOS), shows nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity. In this work, we detected the presence of NADPH-d activity in P. primaurelia. We characterized this activity histochemically by examining its specificity for β-NADPH and α-NADH co-substrates, and sensitivity both to variations in chemico-physical parameters and to inhibitors of enzymes showing NADPH-d activity. Molecules immunologically related to NOS were recognized by the anti-rat brain NOS (bNOS) antibody. Moreover, bNOS immunoreactivity and NADPH-d activity sites were found to be co-localized. The non-denaturing electrophoresis, followed by exposure to β-NADPH or α-NADH co-substrates, revealed the presence of a band of apparent molecular mass of about 124 kDa or a band of apparent molecular mass of about 175 kDa, respectively. In immunoblot experiments, the bNOS antibody recognized a single band of apparent molecular mass of about 123 kDa.

Original languageEnglish
Pages (from-to)201-208
Number of pages8
JournalEuropean Journal of Protistology
Volume42
Issue number3
DOIs
Publication statusPublished - Sep 29 2006

Fingerprint

Paramecium
NADPH Dehydrogenase
nitric oxide synthase
NADP
NADP (coenzyme)
NAD(P)H dehydrogenase (quinone)
NAD (coenzyme)
molecular weight
Nitric Oxide Synthase
NAD
nitric oxide
Nitric Oxide
brain
antibodies
Antibodies
enzyme inhibitors
cholinergic agents
Brain
Enzyme Inhibitors
acetylcholine

Keywords

  • Ciliates
  • NADPH-diaphorase activity
  • Nitric oxide synthase immunoreactivity
  • Paramecium

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Agricultural and Biological Sciences (miscellaneous)
  • Microbiology

Cite this

Amaroli, A., Ognibene, M., Trielli, F., Trombino, S., Falugi, C., & Delmonte Corrado, M. U. (2006). Detection of NADPH-diaphorase activity in Paramecium primaurelia. European Journal of Protistology, 42(3), 201-208. https://doi.org/10.1016/j.ejop.2006.05.002

Detection of NADPH-diaphorase activity in Paramecium primaurelia. / Amaroli, Andrea; Ognibene, Marzia; Trielli, Francesca; Trombino, Sonya; Falugi, Carla; Delmonte Corrado, Maria Umberta.

In: European Journal of Protistology, Vol. 42, No. 3, 29.09.2006, p. 201-208.

Research output: Contribution to journalArticle

Amaroli, A, Ognibene, M, Trielli, F, Trombino, S, Falugi, C & Delmonte Corrado, MU 2006, 'Detection of NADPH-diaphorase activity in Paramecium primaurelia', European Journal of Protistology, vol. 42, no. 3, pp. 201-208. https://doi.org/10.1016/j.ejop.2006.05.002
Amaroli, Andrea ; Ognibene, Marzia ; Trielli, Francesca ; Trombino, Sonya ; Falugi, Carla ; Delmonte Corrado, Maria Umberta. / Detection of NADPH-diaphorase activity in Paramecium primaurelia. In: European Journal of Protistology. 2006 ; Vol. 42, No. 3. pp. 201-208.
@article{b6f2bccd93134a43901acc98e02cfc43,
title = "Detection of NADPH-diaphorase activity in Paramecium primaurelia",
abstract = "Recently, we showed that Paramecium primaurelia synthesizes molecules functionally related to the cholinergic system and involved in modulating cell-cell interactions leading to the sexual process of conjugation. It is known that nitric oxide (NO) plays a role in regulating the release of transmitter molecules, such as acetylcholine, and that the NO biosynthetic enzyme, nitric oxide synthase (NOS), shows nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity. In this work, we detected the presence of NADPH-d activity in P. primaurelia. We characterized this activity histochemically by examining its specificity for β-NADPH and α-NADH co-substrates, and sensitivity both to variations in chemico-physical parameters and to inhibitors of enzymes showing NADPH-d activity. Molecules immunologically related to NOS were recognized by the anti-rat brain NOS (bNOS) antibody. Moreover, bNOS immunoreactivity and NADPH-d activity sites were found to be co-localized. The non-denaturing electrophoresis, followed by exposure to β-NADPH or α-NADH co-substrates, revealed the presence of a band of apparent molecular mass of about 124 kDa or a band of apparent molecular mass of about 175 kDa, respectively. In immunoblot experiments, the bNOS antibody recognized a single band of apparent molecular mass of about 123 kDa.",
keywords = "Ciliates, NADPH-diaphorase activity, Nitric oxide synthase immunoreactivity, Paramecium",
author = "Andrea Amaroli and Marzia Ognibene and Francesca Trielli and Sonya Trombino and Carla Falugi and {Delmonte Corrado}, {Maria Umberta}",
year = "2006",
month = "9",
day = "29",
doi = "10.1016/j.ejop.2006.05.002",
language = "English",
volume = "42",
pages = "201--208",
journal = "European Journal of Protistology",
issn = "0932-4739",
publisher = "Urban und Fischer Verlag GmbH und Co. KG",
number = "3",

}

TY - JOUR

T1 - Detection of NADPH-diaphorase activity in Paramecium primaurelia

AU - Amaroli, Andrea

AU - Ognibene, Marzia

AU - Trielli, Francesca

AU - Trombino, Sonya

AU - Falugi, Carla

AU - Delmonte Corrado, Maria Umberta

PY - 2006/9/29

Y1 - 2006/9/29

N2 - Recently, we showed that Paramecium primaurelia synthesizes molecules functionally related to the cholinergic system and involved in modulating cell-cell interactions leading to the sexual process of conjugation. It is known that nitric oxide (NO) plays a role in regulating the release of transmitter molecules, such as acetylcholine, and that the NO biosynthetic enzyme, nitric oxide synthase (NOS), shows nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity. In this work, we detected the presence of NADPH-d activity in P. primaurelia. We characterized this activity histochemically by examining its specificity for β-NADPH and α-NADH co-substrates, and sensitivity both to variations in chemico-physical parameters and to inhibitors of enzymes showing NADPH-d activity. Molecules immunologically related to NOS were recognized by the anti-rat brain NOS (bNOS) antibody. Moreover, bNOS immunoreactivity and NADPH-d activity sites were found to be co-localized. The non-denaturing electrophoresis, followed by exposure to β-NADPH or α-NADH co-substrates, revealed the presence of a band of apparent molecular mass of about 124 kDa or a band of apparent molecular mass of about 175 kDa, respectively. In immunoblot experiments, the bNOS antibody recognized a single band of apparent molecular mass of about 123 kDa.

AB - Recently, we showed that Paramecium primaurelia synthesizes molecules functionally related to the cholinergic system and involved in modulating cell-cell interactions leading to the sexual process of conjugation. It is known that nitric oxide (NO) plays a role in regulating the release of transmitter molecules, such as acetylcholine, and that the NO biosynthetic enzyme, nitric oxide synthase (NOS), shows nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity. In this work, we detected the presence of NADPH-d activity in P. primaurelia. We characterized this activity histochemically by examining its specificity for β-NADPH and α-NADH co-substrates, and sensitivity both to variations in chemico-physical parameters and to inhibitors of enzymes showing NADPH-d activity. Molecules immunologically related to NOS were recognized by the anti-rat brain NOS (bNOS) antibody. Moreover, bNOS immunoreactivity and NADPH-d activity sites were found to be co-localized. The non-denaturing electrophoresis, followed by exposure to β-NADPH or α-NADH co-substrates, revealed the presence of a band of apparent molecular mass of about 124 kDa or a band of apparent molecular mass of about 175 kDa, respectively. In immunoblot experiments, the bNOS antibody recognized a single band of apparent molecular mass of about 123 kDa.

KW - Ciliates

KW - NADPH-diaphorase activity

KW - Nitric oxide synthase immunoreactivity

KW - Paramecium

UR - http://www.scopus.com/inward/record.url?scp=33748324871&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748324871&partnerID=8YFLogxK

U2 - 10.1016/j.ejop.2006.05.002

DO - 10.1016/j.ejop.2006.05.002

M3 - Article

VL - 42

SP - 201

EP - 208

JO - European Journal of Protistology

JF - European Journal of Protistology

SN - 0932-4739

IS - 3

ER -