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 language | English |
|---|---|
| Pages (from-to) | 201-208 |
| Number of pages | 8 |
| Journal | European Journal of Protistology |
| Volume | 42 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Sep 29 2006 |
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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
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 journal › Article
}
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 -