Hyaluronidase binds differently DPPC, DPPS or GlcNAc-bearing glycolipid biomimetic monolayers.
Bovine testis hyaluronidase (btHyal) had been shown to have direct effects on cancer cells and to be a useful adjuvant in several medicines. Furthermore this enzyme had been found to be membrane-associated. Thus, in this work, the interactions between btHyal and membranes were analyzed by using lipid monolayers at the air-water interface as a biomimetic membrane system. This allowed us to define the btHyal interactions with two residues of hyaluronic acid (a btHyal substrate), GlcNAc and carboxylic group, which are present in cholesteryl-triethoxy-N-acetylglucosamine (Chol-E3-GlcNAc) and in DPPS, respectively. btHyal bound preferentially Chol-E3-GlcNAc monolayers and showed a decreasing affinity for Chol-E3-GlcNAc-DPPC monolayers containing decreasing amount of glycolipid, suggesting a crucial role of the glycolipid GlcNAc. Furthermore the significant btHyal binding to DPPS was not affected by the presence of free GlcNAc in the subphase. These results and the absence of significant binding of btHyal to pure DPPC monolayer suggest that the protein interacts with the lipid monolayer by mimicking the enzyme-substrate interactions or by electrostatic interactions.
Belem-Gonçalves
Silvia
S
Université de Lyon, Villeurbanne F-69622, France.
Matar
Gladys
G
Tsan
Pascale
P
Lafont
Dominique
D
Boullanger
Paul
P
Salim
Vera M
VM
Alves
Tito L M
TL
Lancelin
Jean-Marc
JM
Besson
Françoise
F
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000378:metabolism*; D000117:Acetylglucosamine / Q000737:chemistry / Q000378:metabolism*; D000327:Adsorption; D000818:Animals; D040761:Biomimetic Materials / Q000378:metabolism*; D002021:Buffers; D002417:Cattle; D002784:Cholesterol / Q000737:chemistry; D055119:Elastic Modulus; D006017:Glycolipids / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008853:Microscopy; D008958:Models, Molecular; D010718:Phosphatidylserines / Q000378:metabolism*; D055672:Static Electricity; D013696:Temperature
D002021:Buffers; D006017:Glycolipids; D010718:Phosphatidylserines; D015060:1,2-Dipalmitoylphosphatidylcholine; C038694:dipalmitoylphosphatidylserine; D002784:Cholesterol; D006821:Hyaluronoglucosaminidase; D000117:Acetylglucosamine