Radiation Protection: Radiation Toxicology: neuraminidase enzymes, neuraminidase inhibitors.

1. RADIATION PROTECTION:
RADIATION TOXICOLOGY:
NEURAMINIDASE ENZYMES,
NEURAMINIDASE INHIBITORS.
DMITRI POPOV. MD (RUSSIA), PHD RADIOBIOLOGY.
ADVANCED MEDICAL TECHNOLOGY AND SYSTEMS INC. CANADA.

2. RADIATION TOXICOLOGY.
• Radiation Protection: Radiation Toxicology: neuraminidase
enzymes, neuraminidase inhibitors.
• Dmitri Popov
• Full-text Research Proposal · Feb 2016
• Add resources
• File name: RadiationToxicologyNeuroamidaze.pptx
DOI: 10.13140/RG.2.2.14393.36962

3. NEURAMINIDASE.
HTTPS://EN.WIKIPEDIA.ORG/WIKI/
NEURAMINIDASE
• Neuraminidase enzymes are glycoside hydrolase enzymes (EC 3.2.1.18) that
cleave the glycosidic linkages of neuraminic acids. Neuraminidase enzymes
are a large family, found in a range of organisms. The best-known
neuraminidase is the viral neuraminidase, a drug target for the prevention of
the spread of influenza infection. The viral neuraminidases are frequently
used as antigenic determinants found on the surface of the Influenza virus.
Some variants of the influenza neuraminidase confer more virulence to the
virus than others. Other homologs are found in mammalian cells, which
have a range of functions.
• At least four mammalian sialidase homologs have been described in
the human genome (see NEU1, NEU2, NEU3, NEU4).

4. NEURAMINIDASE.
• There are two major classes of Neuraminidase that cleave exo
or endo poly-sialic acids:
• Exo hydrolysis of α-(2→3)-, α-(2→6)-, α-(2→8)-glycosidic
linkages of terminal sialic acid residues.
• Endo hydrolysis of (2→8)-α-sialosyl linkages in oligo- or
poly(sialic) acids[2]

5. NEURAMINIDASE.
• Schauer R (1982). "Chemistry, metabolism, and biological
functions of sialic acids". Adv Carbohydr Chem Biochem.
Advances in Carbohydrate Chemistry and Biochemistry. 40:
131–234. doi:10.1016/S0065-2318(08)60109-2. ISBN 978-0-
12-007240-8.PMID 6762816.

6. NEURAMINIDASE
• Cabezas JA (August 1991). "Some questions and suggestions
on the type references of the official nomenclature (IUB) for
sialidase(s) and endosialidase". Biochem. J. 278 (Pt 1): 311–
2. doi:10.1042/bj2780311

7. NEURAMINIDAZE
• Sialidase 1 (lysosomal sialidase), also known as NEU1 is a
mammalian lysosomal neuraminidase enzyme which in humans
is encoded by the NEU1 gene.

8. NEURAMINIDASE
• The protein encoded by this gene encodes the lysosomal
enzyme, which cleaves terminal sialic acid residues from
substrates such as glycoproteins and glycolipids. In
the lysosome, this enzyme is part of a heterotrimeric complex
together with beta-galactosidase and cathepsin A (the latter
also referred to as 'protective protein'). Mutations in this gene
can lead tosialidosis.[3]

9. • Sialidase-2 is an enzyme that in humans is encoded by
the NEU2 gene.
• This gene belongs to a family of glycohydrolytic enzymes which
remove sialic acid residues from glycoproteins and glycolipids.
Expression studies in COS7 cells confirmed that this gene
encodes a functional sialidase. Its cytosolic localization was
demonstrated by cell fractionation experiments.

10. NEURAMINIDASE.
• Sialidase-3 is an enzyme that in humans is encoded by
the NEU3 gene.
• This gene product belongs to a family of glycohydrolytic
enzymes which remove sialic acid residues from glycoproteins
and glycolipids. It is localized in the plasma membrane, and its
activity is specific for gangliosides. It may play a role in
modulating the ganglioside content of the lipid bilayer.

11. NEURAMINIDASE
• This gene belongs to a family of glycohydrolytic enzymes which
remove sialic acid residues from glycoproteins and glycolipids.
• Sialidase-4 is an enzyme that in humans is encoded by
the NEU4 gene.

12. VIRAL NEURAMINIDASE
• Viral neuraminidase is a type of neuraminidase found on the surface
of influenza viruses that enables the virus to be released from the host cell.
Neuraminidases are enzymes that cleave sialic acid groups
from glycoproteins and are required for influenza virus replication.
• When influenza virus replicates, it attaches to the interior cell surface
using hemagglutinin, a molecule found on the surface of the virus that binds
to sialic acid groups. Sialic acids are found on various glycoproteins at the
host cell surface, and the virus exploits these groups to bind the host cell. In
order for the virus to be released from the cell, neuraminidase must
enzymatically cleave the sialic acid groups from host glycoproteins

13. VIRAL NEURAMINIDASE
• Since the cleavage of the sialic groups is an integral part of
influenza replication, blocking the function of neuraminidase
with neuraminidase inhibitors is an effective way to treat
influenza.
• A single hemagglutinin-neuraminidase protein can combine
neuraminidase and hemagglutinin functions, such as in mumps
virus and human parainfluenza virus.

14. VIRAL NEURAMINIDASE.
• Hemagglutinin or haemagglutinin (British English) refers to a
substance that causes red blood cells (RBCs) to agglutinate.
This process is called hemagglutination or haemagglutination.
• Antibodies[1] and lectins[2] are commonly known
hemagglutinins.

15. BACTERIAL NEURAMINIDASE
• Bacterial neuraminidase is type of neuraminidase and a
virulence factor for many bacteria including Bacteroides
fragilis and Pseudomonas aeruginosa. Its function is to cleave a
sialic acid residue off ganglioside-GM1 (a modulator of cell
surface and receptor activity) turning it into asialo-GM1 to
which type 4 pilli (attachment factors) bind preferentially.
• https://en.wikipedia.org/wiki/Bacterial_neuraminidase

16. BACTERIAL NEURAMINIDASE
• Many respiratory pathogens, including Hemophilus influenzae,
Streptococcus pneumoniae, and Pseudomonas aeruginosa,
express neuraminidases that can cleave alpha2,3-linked sialic
acids from glycoconjugates. As mucosal surfaces are heavily
sialylated, neuraminidases have been thought to modify
epithelial cells by exposing potential bacterial receptors.

17. BACTERIAL NEURAMINIDASE.
• J Clin Invest. 2006 Aug;116(8):2297-2305.
• Bacterial neuraminidase facilitates mucosal infection by
participating in biofilm production.
• Soong G1, Muir A, Gomez MI, Waks J, Reddy B, Planet P, Singh
PK, Kaneko Y, Wolfgang MC, Hsiao YS, Tong L, Prince A.

18. BACTERIAL NEURAMINIDASES.
• Front Med (Lausanne). 2015; 2: 14.
• Published online 2015 Mar 17. doi: 10.3389/fmed.2015.00014
• PMCID: PMC4362343
• Neuroinflammation Induced by Intracerebroventricular Injection of Microbial
Neuraminidase
• Pablo Granados-Durán,1 María D. López-Ávalos,1 Jesús M. Grondona,1 María
del Carmen Gómez-Roldán,1Manuel Cifuentes,1,2 Margarita Pérez-
Martín,1 Martina Alvarez,3 Fernando Rodríguez de Fonseca,4 and Pedro
Fernández-Llebrez1,*

19. BACTERIAL NEURAMINIDASES
• Kelly and Greiff demonstrated in mice the high toxic effects of
bacterial and viral NA when injected directly into the brain.
Also, in humans, NA activity seems to be associated with
adverse prognosis in pneumococcal meningitis

20. BACTERIAL NEURAMINIDASES
• Kelly RT, Greiff D. Neuraminidase and neuraminidase-labile
substrates in experimental influenza virus encephalitis. Biochim
Biophys Acta (1965) 110:548–5310.1016/S0926-
6593(65)80068-6 [Cross Ref]
• 12. Kelly R, Greiff D. Toxicity of pneumococcal
neuraminidase. Infect Immun (1970) 2:115–7.

21. BACTERIAL NEURAMINIDASES.
• O’Toole RD, Goode L, Howe C.. Neuraminidase activity in
bacterial meningitis. J Clin Invest (1971)50:979–
85.10.1172/JCI106591
• 14. Löve A, Rydbeck R, Kristensson K, Orvell C, Norrby
E.. Hemagglutinin-neuraminidase glycoprotein as a
determinant of pathogenicity in mumps virus hamster
encephalitis: analysis of mutants selected with monoclonal
antibodies. J Virol (1985) 53:67–74.

22. BACTERIAL NEURAMINIDASES.
• Finsterer J, Hess B.. Neuromuscular and central nervous system
manifestations of Clostridium
perfringens infections. Infection (2007) 35:396–
405.10.1007/s15010-007-6345-z

23. BACTERIAL NEURAMINIDASES.
• Kelly RT, Greiff D. Neuraminidase and neuraminidase-labile
substrates in experimental influenza virus encephalitis. Biochim
Biophys Acta (1965) 110:548–5310.1016/S0926-
6593(65)80068-6 [Cross Ref]

24. NEURAMINIDASES AND IMMUNOTHERAPY
OF TUMORS.
• The effect of active immunotherapy with Vibrio cholerae neuraminidase-treated syngeneic
tumor cells (VCN-cells) following radiotherapy has been studied with 3-
methylcholanthrene-induced fibrosarcoma, M-79, transplanted to the thigh of C3H/HeJ
mice. When the tumors reached 4-8 mm in diameter, various treatments were started. X-
irradiation with 2000 rad in a single dose induced a complete regression of 24 out of 103
tumors (23.3%). The inoculation of 1× 106of VCN-cells to the tumor-bearing animals,
every other day for a total of three doses, caused a complete regression of 6 out of 57
tumors (10.5%). Treatments of animals with the immunotherapy starting 1 day after X-
irradiation of tumors with 2000 rad resulted in a complete regression of 22 out of 58
tumors (37.9%). The median survival time of animals that received combined radiotherapy
and immunotherapy was longer than that observed after either treatment alone.
• Copyright 1975 Academic Press, Inc.

25. NEURAMINIDASES AND IMMUNOTHERAPY
OF TUMORS.
• Chang W. Song and Seymour H. Levitt (1975) Immunotherapy
with Neuraminidase-Treated Tumor Cells after Radiotherapy.
Radiation Research: December 1975, Vol. 64, No. 3, pp. 485-
491.
• Immunotherapy with Neuraminidase-Treated Tumor Cells after
Radiotherapy
• Chang W. Song and Seymour H. Levitt

26. IMMUNOTHERAPY
• Science. 1971 Nov 5;174(4009):591-3.
• Immunotherapy of cancer: immunospecific rejection of tumors in recipients of
neuraminidase-treated tumor cells plus BCG.
• Simmons RL, Rios A.
• Abstract
• Firmly established methylcholanthrene fibrosarcomas in syngeneic mice will totally
disappear if the hosts are treated with living tumor cells that have been exposed to Vibrio
cholerae neuraminidase in vitro. The effect is magnified by the simultanieous injection of
a nonspecific immunostimulant, BCG. The rejection of the methylcholanthrene tumor is
immunospecific and can be induced only with tumor cells, treated with Vibrio cholerae
neuraminidase, identical in type with the growing tumor.

27. NEURAMINIDASES.
• Biochemistry. 1984 Mar 27;23(7):1442-8.
• Modulation of neuraminidase activity by the physical state of
phospholipid bilayers containing gangliosides Gd1a and Gt1b.
• Myers M, Wortman C, Freire E.

28. HUMAN NEURAMINIDASES.
• Lysosomal neuraminidase is present In virtually all vertebrate
tissues and cell types and has been purified and characterized
from many sources, including placenta, mammary gland, brain,
kidney, liver, testis, thyroid, salivary gland, leukocytes,
lymphocytes, macrophages and fibroblasts

29. RADIATION AND GLYCOSYLATION.
• Biochem Biophys Res Commun. 1997 Nov 17;240(2):395-8.
• Altered N-glycosylation of glucose transporter-1 associated
with radiation-induced tumorigenesis of human cell hybrids.
• Noto Y1, Iwazaki A, Nagao J, Sumiyama Y, Redpath
JL, Stanbridge EJ, Kitagawa T.

30. RADIATION AND GLYCOSYLATION.
• “Studies on human cell hybrids between a cervical carcinoma cell line,
HeLa, and normal fibroblasts have indicated that their tumorigenicity
is under the control of a putative tumor suppressor on chromosome
11. We have previously demonstrated that a tumorigenic cell hybrid
CGL4 expresses a larger glucose transporter, GLUT1, due to altered
glycosylation when compared to the nontumorigenic counterpart
CGL1. In this study, we demonstrated this glycosylation change in
GLUT1 in gamma-ray-induced tumorigenic mutants (GIMs) isolated
from CGL1 cells as expressing a tumor-associated surface antigen,
intestinal alkaline phosphatase.”

31. RADIATION AND GLYCOSYLATION.
• “In contrast, GLUT1 in the gamma-irradiated nontumorigenic
control cells (CONs) did not show this alteration. In accordance
with this glycosylation change, affinity to 2-deoxyglucose in
these GIM clones was increased by about twofold when
compared to the nontumorigenic CONs. These results suggest
a close correlation between the glycosylation change in GLUT1
with increased affinity to D-glucose and tumorigenicity of these
human cell hybrids.”

32. RADIATION AND GLYCOSYLATION.
• J Pharm Biomed Anal. 2016 Jan 25;118:380-6. doi: 10.1016/j.jpba.2015.11.010. Epub 2015 Nov 14.
• Changes of protein glycosylation in the course of radiotherapy.
• Tóth E1, Vékey K2, Ozohanics O3, Jekő A3, Dominczyk I4, Widlak P4, Drahos L5.
• Author information
• Abstract
• This is the first study of changes in protein glycosylation due to exposure of human subjects to
ionizing radiation. Site specific glycosylation patterns of 7 major plasma proteins were analyzed; 171
glycoforms were identified; and the abundance of 99 of these was followed in the course of cancer
radiotherapy in 10 individual patients. It was found that glycosylation of plasma proteins does change
in response to partial body irradiation (∼ 60 Gy), and the effects last during follow-up; the abundance
of some glycoforms changed more than twofold. Both the degree of changes and their time-evolution
showed large inter-individual variability.

33. INHIBITION OF NEURAMINIDASES.
• “Inhibition of neuraminidase (NA) activity prevents release of progeny
virions from influenza-infected cells and removal of neuraminic
(sialic) acid moieties from glycans attached to hemagglutinin (HA).
Neuraminic acid moieties situated near the HA receptor-binding site
can reduce the efficiency of virus binding and decrease viral
dependence on NA activity for replication. With the use of reverse
genetics technique, we investigated the effect of glycans attached at
Asn 94a, 129, and 163 on the virus susceptibility to NA inhibitors in
MDCK cells and demonstrated that the glycan attached at Asn 163
plays a dominant role in compensation for the loss of NA activity.”

34. INHIBITION OF NEURAMINIDASES
• Effect of Hemagglutinin Glycosylation on Influenza Virus
Susceptibility to Neuraminidase Inhibitors
• Vasiliy P. Mishin1,†,
• Dmitri Novikov1,‡,
• Frederick G. Hayden1,2, and
• Larisa V. Gubareva1,*

35. NEURAMINIDASES.
• The two surface glycoproteins, hemagglutinin (HA) and
neuraminidase (NA), of influenza A virus interact with cellular
receptors containing terminal neuraminic acid (NeuAc)
moieties. HA initiates infection by binding to cellular receptors,
whereas NA destroys the receptors by cleaving off NeuAc
moieties

36. NEURAMIDASES INHIBITION
• JOURNAL OF VIROLOGY, Oct. 2005, p. 12416–12424 Vol. 79, No. 19
0022-538X/05/$08.000 doi:10.1128/JVI.79.19.12416–12424.2005
Copyright © 2005, American Society for Microbiology. All Rights
Reserved. Effect of Hemagglutinin Glycosylation on Influenza Virus
Susceptibility to Neuraminidase Inhibitors Vasiliy P. Mishin,1 † Dmitri
Novikov,1 ‡ Frederick G. Hayden,1,2 and Larisa V. Gubareva1 *
Department of Internal Medicine, School of Medicine, University of
Virginia, Charlottesville, Virginia1 and Department of Pathology,
School of Medicine, University of Virginia, Charlottesville, Virginia2

37. NEURAMINIC ACID.
• Neuraminic acid (5-amino-3,5-dideoxy-D-glycero-D-galacto-non-2-ulosonic
acid) is a 9-carbon monosaccharide (a nonose), a derivative of a ketononose.
Neuraminic acid may be visualized as the product of an aldol-
condensation product of pyruvic acid and D-mannosamine (2-amino-2-deoxy-
mannose). Neuraminic acid does not occur naturally, but many of its derivatives
are found widely distributed in animal tissues and in bacteria, especially
in glycoproteins and gangliosides. The N- or O-substituted derivatives of
neuraminic acid are collectively known as sialic acids, the predominant form in
mammalian cells being N-acetylneuraminic acid. The amino group bears either
an acetyl or a glycolyl group. The hydroxyl substituents may vary considerably:
acetyl, lactyl, methyl, sulfate and phosphate groups have been found.

38. NEURAMINIC ACID.
• Br J Pharmacol. 1978 Oct;64(2):301-4.
• Inhibition by N-acetyl neuraminic acid of platelet thrombogenesis induced by laser injury to rat and
hamster venules.
• Born GV, Kovács IB.
• Abstract
• 1 In rats and hamsters under barbiturate anaesthesia, laser radiation to venules about 50 micrometer
in diameter in mesoappendix and cheek pouch respectively caused the formation of platelet thrombi
which occluded the vessels in about 9 min. 2 This occlusion time was significantly prolonged by the
intravenous injection of N-acetyl neuraminic acid (NANA) but not by D-glucuronic acid or beta-
methoxyneuraminic acid, in doses which had no effect on blood pH or on the condition of the
animals. 3 The results confirm the anti-thrombotic effect of NANA previously demonstrated with
another technique.

39. CYTOTOXICITY
• The antibody-dependent cell-mediated cytotoxicity (ADCC) is a
mechanism of cell-mediated immune defense whereby an
effector cell of the immune system actively lyses a target cell,
whose membrane-surface antigens have been bound by
specific antibodies. In humans, ADCC is usually mediated by
IgG.[citation needed] It is one of the mechanisms through which
antibodies, as part of the humoral immune response, can act to
limit and contain infection

40. CYTOTOXICITY, HEMAGGLUTININS,
NEURAMINIDASES.
• Detection of neutralizing antibodies (nAbs) to influenza A virus
hemagglutinin (HA) antigens by conventional serological assays is
currently the main immune correlate of protection for influenza
vaccines However, current prepandemic avian influenza vaccines are
poorly immunogenic in inducing nAbs despite considerable
protection conferred. Recent studies show that Ab-dependent cell-
mediated cytotoxicity (ADCC) to HA antigens are readily detectable in
the sera of healthy individuals and patients with influenza infection

41. CYTOTOXICITY
• The typical ADCC involves activation of NK cells by antibodies. A NK cell expresses Fc
receptors, mostly CD16. These receptors recognize, and bind to, the Fc portion of an
antibody, such as IgG, which has bound to the surface of a pathogen-infected target cell.
The most common Fc receptor on the surface of an NK cell is called CD16 or FcγRIII. Once
the Fc receptor binds to the Fc region of IgG, the Natural Killer cell
releases cytokines such as IFN-γ
• During replication of a virus some of the viral proteins are expressed on the cell surface
membrane of the infected cell. Antibodies can then bind to these viral proteins. Next, the
NK cells which have Fc Receptors will bind to that antibody, inducing the NK cell to
release proteins such as perforin and proteases known as granzymes, which causes the
lysis of the infected cell to hinder the spread of the virus.
• Furthermore, NK cells are involved in killing tumor cells and other cells that may lack MHC
I on their surface, indicating a non-self cell. This is because, generally, all nucleated cells
(which excludes RBCs) of the body contain MHC I.

42. HUMAN NEURAMINIDASES AND ACUTE
RADIATION SYNDROMES.
• Inhibiting human neuraminidases before and after irradiation
could provide a novel mechanism to prevent Acute Radiation
Syndromes.

43. LITERATURE.
• https://en.wikipedia.org/wiki/Antibody-dependent_cell-
mediated_cytotoxicity
• http://www.jbc.org/content/275/48/37657.full
• https://books.google.ca/books?id=2IXMBQAAQBAJ&pg=PA131&lpg=
PA131&dq=peroxidases+and+lysosomal+activation&source=bl&ots
=VZxbVXtMfM&sig=TO2JSWMpz1ZTl8Xxi_xnhFQRofg&hl=en&sa=X&v
ed=0ahUKEwigtuPQi97PAhVI7SYKHdgJC-
0Q6AEIRDAH#v=onepage&q=peroxidases%20and%20lysosomal%20ac
tivation&f=false

44. LITERATURE.
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4362343/
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4943536/
• https://www.ncbi.nlm.nih.gov/pubmed/26609677