title: Activity of ulilysin, an archaeal PAPP-A-related gelatinase and IGFBP protease.
abstract: Human growth and development are conditioned by insulin-like growth factors (IGFs), which have also implications in pathology. Most IGF molecules are sequestered by IGF-binding proteins (IGFBPs) so that exertion of IGF activity requires disturbance of these complexes. This is achieved by proteolysis mediated by IGFBP proteases, among which the best characterised is human PAPP-A, the first member of the pappalysin family of metzincins. We have previously identified and studied the only archaeal homologue found to date, Methanosarcina acetivorans ulilysin. This is a proteolytically functional enzyme encompassing a pappalysin catalytic domain and a pro-domain involved in maintenance of latency of the zymogen, proulilysin. Once activated, the protein hydrolyses IGFBP-2 to -6 and insulin chain beta in vitro. We report here that ulilysin is also active against several other substrates, viz (azo)casein, azoalbumin, and extracellular matrix components. Ulilysin has gelatinolytic but not collagenolytic activity. Moreover, the proteolysis-resistant skeletal proteins actin and elastin are also cleaved, as is fibrinogen, but not plasmin and alpha1-antitrypsin from the blood coagulation cascade. Ulilysin develops optimal activity at pH 7.5 and strictly requires peptide bonds preceding an arginine residue, as determined by means of a novel fluorescence resonance energy transfer assay, thus pointing to biotechnological applications as an enzyme complementary to trypsin.
authors.affiliation: Departament de Biologia Estructural, Institut de Biologia Molecular de Barcelona, CSIC, c/ Jordi Girona, 18-26, E-08034, Barcelona, Spain.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Tallant
authors.forename: Raquel
authors.initials: R
authors.lastname: García-Castellanos
authors.forename: Aniebrys
authors.initials: A
authors.lastname: Marrero
authors.forename: Francesc
authors.initials: F
authors.lastname: Canals
authors.forename: Yongzheng
authors.initials: Y
authors.lastname: Yang
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Reymond
authors.forename: Maria
authors.initials: M
authors.lastname: Solà
authors.forename: Ulrich
authors.initials: U
authors.lastname: Baumann
authors.forename: F Xavier
authors.initials: FX
authors.lastname: Gomis-Rüth
mesh_terms: D001105:Archaea / Q000201:enzymology*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004789:Enzyme Activation; D018093:Gelatinases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D045726:Metalloproteases / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D045726:Metalloproteases; D010447:Peptide Hydrolases; C512490:ulilysin, Methanosarcina acetivorans; D018093:Gelatinases