A Diverse Range of Bacterial and Eukaryotic Chitinases Hydrolyzes the LacNAc (Galβ1-4GlcNAc) and LacdiNAc (GalNAcβ1-4GlcNAc) Motifs Found on Vertebrate and Insect Cells

Rikki F. Frederiksen, Yayoi Yoshimura, Birgit G. Storgaard, Dafni K. Paspaliari, Bent O. Petersen, Kowa Chen, Tanja Larsen, Jens Øllgaard Duus, Hanne Ingmer, Nicolai V. Bovin, Ulrika Westerlind, Ola Blixt, Monica M. Palcic, Jorgen J. Leisner

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

There is emerging evidence that chitinases have additional functions beyond degrading environmental chitin, such as involvement in innate and acquired immune responses, tissue remodeling, fibrosis, and serving as virulence factors of bacterial pathogens. We have recently shown that both the human chitotriosidase and a chitinase from Salmonella enterica serovar Typhimurium hydrolyze LacNAc from Gal beta 1-4GlcNAc beta-tetramethylrhodamine (LacNAc-TMR (Gal beta 1-4GlcNAc beta (CH2)(8)-CONH(CH2)(2)NHCO-TMR)), a fluorescently labeled model substrate for glycans found in mammals. In this study we have examined the binding affinities of the Salmonella chitinase by carbohydrate microarray screening and found that it binds to a range of compounds, including five that contain LacNAc structures. We have further examined the hydrolytic specificity of this enzyme and chitinases from Sodalis glossinidius and Polysphondylium pallidum, which are phylogenetically related to the Salmonella chitinase, as well as unrelated chitinases from Listeria monocytogenes using the fluorescently labeled substrate analogs LacdiNAc-TMR (GalNAc beta 1-4GlcNAc beta-TMR), LacNAc-TMR, and LacNAc beta 1-6LacNAc beta-TMR. We found that all chitinases examined hydrolyzed LacdiNAc from the TMR aglycone to various degrees, whereas they were less active toward LacNAc-TMR conjugates. LacdiNAc is found in the mammalian glycome and is a common motif in invertebrate glycans. This substrate specificity was evident for chitinases of different phylogenetic origins. Three of the chitinases also hydrolyzed the beta 1-6 bond in LacNAc beta 1-6LacNAc beta-TMR, an activity that is of potential importance in relation to mammalian glycans. The enzymatic affinities for these mammalian-like structures suggest additional functional roles of chitinases beyond chitin hydrolysis.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume590
Issue number9
Pages (from-to)5354-5366
Number of pages13
ISSN0021-9258
DOIs
Publication statusPublished - 2015
Externally publishedYes

Keywords

  • Phylogenetic origin
  • Arthropoda Invertebrata Animalia (Animals, Arthropods, Insects, Invertebrates) - Insecta [75300] insect common
  • Chordata Animalia (Animals, Chordates, Nonhuman Vertebrates, Vertebrates) - Vertebrata [85150] Vertebrata higher_taxa
  • Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Regular Nonsporing Gram-Positive Rods [07830] Listeria monocytogenes species
  • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Salmonella enterica species serovar-Typhimurium hydrolyze LacNAc Sodalis glossinidius species strain-morsitans SgChiA SG1474
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Myxophyta [15700] Polysphondylium pallidum species
  • chitin 1398-61-4
  • chitinase 9001-06-3 EC 3.2.1.14
  • Gal-beta-1-4GlcNAc LacNAc 32181-59-2
  • GalNAc-bata-1 4GlcNAc LacdiNAc
  • 10068, Biochemistry studies - Carbohydrates
  • 10802, Enzymes - General and comparative studies: coenzymes
  • 31000, Physiology and biochemistry of bacteria
  • 51518, Plant physiology - Enzymes
  • 64076, Invertebrata: comparative, experimental morphology, physiology and pathology - Insecta: physiology
  • Biochemistry and Molecular Biophysics
  • carbohydrate microarray screeningc laboratory techniques
  • Enzymology
  • Biochemistry
  • Flurorescently labeled oligosaccharides
  • Salmonella-enterica
  • Plant chitinases
  • Binding-proteins
  • Repeating unit
  • Chitotriosidase
  • Transmission
  • Separation
  • Expression
  • Infection
  • Binding energy
  • Chitin
  • Hydrolysis
  • Mammals
  • Pathogens
  • Polysaccharides
  • Bacterial pathogens
  • Binding affinities
  • Carbohydrate microarrays
  • Labeled substrates
  • Listeria monocytogenes
  • Salmonella enterica serovar Typhimurium
  • Substrate specificity
  • Virulence factors
  • Salmonella

Fingerprint Dive into the research topics of 'A Diverse Range of Bacterial and Eukaryotic Chitinases Hydrolyzes the LacNAc (Galβ1-4GlcNAc) and LacdiNAc (GalNAcβ1-4GlcNAc) Motifs Found on Vertebrate and Insect Cells'. Together they form a unique fingerprint.

Cite this