Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei

Autor: Nicolas Langenegger, Aline Moser, Olivier Felix Biner, Stefan Schürch, Lukas S. Kopp, Johann Schaller, Lucia Kuhn-Nentwig, Christoph von Ballmoos, Urs Kämpfer, Christian Trachsel, Wolfgang Nentwig
Přispěvatelé: University of Zurich
Jazyk: angličtina
Rok vydání: 2015
Předmět:
Glycosylation
Spider Venoms
Molecular Sequence Data
lcsh:Medicine
Hyaluronoglucosaminidase
610 Medicine & health
10071 Functional Genomics Center Zurich
Venom
1100 General Agricultural and Biological Sciences
complex mixtures
Dermatan sulfate
chemistry.chemical_compound
1300 General Biochemistry
Genetics and Molecular Biology
Tandem Mass Spectrometry
Hyaluronidase
Hydrolase
540 Chemistry
medicine
Animals
Amino Acid Sequence
lcsh:Science
chemistry.chemical_classification
1000 Multidisciplinary
Multidisciplinary
Base Sequence
biology
lcsh:R
Spiders
Heparan sulfate
biology.organism_classification
Cupiennius salei
Recombinant Proteins
Enzyme
chemistry
Biochemistry
Spectrometry
Mass
Matrix-Assisted Laser Desorption-Ionization
570 Life sciences
590 Animals (Zoology)
lcsh:Q
Research Article
medicine.drug
Zdroj: PLoS ONE
PLoS ONE, Vol 10, Iss 12, p e0143963 (2015)
PLoS ONE, 10 (12)
Biner, Olivier Felix; Trachsel, Christian; Moser, Aline Isabelle; Kopp, Lukas; Langenegger, Nicolas; Kämpfer, Urs; von Ballmoos, Christoph; Nentwig, Wolfgang; Schürch, Stefan; Schaller, Johann; Kuhn-Nentwig, Lucia Gerda (2015). Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. PLoS ONE, 10(12), e0143963. Public Library of Science 10.1371/journal.pone.0143963
ISSN: 1932-6203
DOI: 10.7892/boris.74980
Popis: Structure of Cupiennius salei venom hyaluronidase Hyaluronidases are important venom components acting as spreading factor of toxic compounds. In several studies this spreading effect was tested on vertebrate tissue. However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. A C-terminal EGF-like domain was identified in spider hyaluronidase using InterPro. The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40–60°C, and 0.2 M KCl. Divalent ions did not enhance HA degradation activity, indicating that they are not recruited for catalysis. Function of venom hyaluronidases Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. The enzyme significantly enhances the neurotoxic activity of CsTx-1. Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques.
PLoS ONE, 10 (12)
ISSN:1932-6203
Databáze: OpenAIRE
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