The Pro-regions of lysyl oxidase and lysyl oxidase-like 1 are required for deposition onto elastic fibers.
These studies were undertaken to determine how lysyl oxidase (LOX) and lysyl oxidase like-1 (LOXL) enzymes are targeted to their substrates in the extracellular matrix. Full-length LOX/LOXL and constructs containing just the pro-regions of each enzyme localized to elastic fibers when expressed in cultured cells. However, the LOXL catalytic domain without the pro-region was secreted into the medium but did not associate with matrix. Ligand blot and mammalian two-hybrid assays confirmed an interaction between tropoelastin and the pro-regions of both LOX and LOXL. Immunofluorescence studies localized both enzymes to elastin at the earliest stages of elastic fiber assembly. Our results showed that the pro-regions of LOX and LOXL play a significant role in directing the deposition of both enzymes onto elastic fibers by mediating interactions with tropoelastin. These findings confirmed that an important element of substrate recognition lies in the pro-domain region of the molecule and that the pro-form of the enzyme is what initially interacts with the matrix substrate. These results have raised the interesting possibility that sequence differences between the pro-domain of LOX and LOXL account for some of the functional differences observed for the two enzymes.
Thomassin
Laetitia
L
Institut de Biologie et Chimie des Protéines, CNRS/Université Lyon 1 UMR-5086, 7 passage du Vercors, 69367 Lyon Cedex 07, France.
Werneck
Claudio C
CC
Broekelmann
Thomas J
TJ
Gleyzal
Claudine
C
Hornstra
Ian K
IK
Mecham
Robert P
RP
Sommer
Pascal
P
D000594:Amino Acid Oxidoreductases / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D020134:Catalytic Domain; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D016296:Mutagenesis; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D008249:Protein-Lysine 6-Oxidase / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D014333:Tropoelastin / Q000737:chemistry; D020798:Two-Hybrid System Techniques
D018076:DNA, Complementary; D008024:Ligands; D011994:Recombinant Proteins; D014333:Tropoelastin; D004247:DNA; D008156:Luciferases; D000594:Amino Acid Oxidoreductases; C445313:LOXL1 protein, human; D008249:Protein-Lysine 6-Oxidase