Structural features associated with the binding of glutamine-containing peptides to Factor XIII

Marinescu, Anca; Cleary, David B.; Littlefield, Tara R; Maurer, Muriel C.

doi:10.1016/s0003-9861(02)00407-1

Cited by 11 publications

(19 citation statements)

References 35 publications

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“…FXIII was nonproteolytically activated under a series of conditions and TGase activity monitored using a modified version of the Dade-Behring Berichrom assay [31]. When incubated in the presence of several different divalent metals including 50 mM Ca 2+ , Mg 2+ , Ba 2+ or Cu 2+ , only Ca 2+ displayed activity (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…FXIII was activated for 10 min at 37 °C before adding 3 μl to the assay. For each assay, the volume of activator reagent (163 μl) and detector reagent (250 μl) as well as the concentration of FXIII (300 nM) and K9 (500 μM) were held constant [31]. The assay contents were placed in a Cary 100 UV/vis spectrophotometer for 2 min at 37 °C for equilibration before the K9 substrate peptide was added.…”

Section: Methodsmentioning

confidence: 99%

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Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Woofter¹,

Maurer²

2011

Archives of Biochemistry and Biophysics

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Factor XIII catalyzes formation of γ-glutamyl-ε-lysyl crosslinks within fibrin clots. FXIII A2 can be activated proteolytically with thrombin and low mM Ca2+ or nonproteolytically with high monovalent/divalent cations along with low mM Ca2+. Physiologically, FXIII A2 is poised to respond to transient influxes of Ca2+ in a Na+ containing environment. A successful strategy to monitor FXIII conformational events is hydrogen-deuterium exchange (HDX) coupled with mass spectrometry. FXIII A2 was examined in the presence of different cations (Ca2+, Mg2+, Ba2+, Cu2+, Na+, TMAC+, and EDA2+) ranging from 1–2 mM, physiological Ca2+ concentration, to 50–500mM for nonproteolytic activation. Increases in FXIII solvent exposure could already be observed at 1 mM Ca2+ for the dimer interface, the catalytic site, and glutamine substrate regions. By contrast, solvent protection was observed at the secondary cleavage site. These events occurred even though 1mM Ca2+ is insufficient for FXIII activation. The metals 1mM Mg2+, 1mM Ba2+, and 1mM Cu2+ each led to conformational changes, many in the same FXIII regions as Ca2+. FXIII could also be activated nonproteolytically with 500mM tetramethylammonium chloride (TMAC+) and 500mM ethylenediamine (EDA2+), both with 2mM Ca2+. These different HDX studies help reveal the first FXIII segments that respond to physiological Ca2+ levels.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Woofter¹,

Maurer²

2011

Archives of Biochemistry and Biophysics

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“…In summary, Asn1 has a major role in binding N1-a 2 PI(1-12) to FXIIIa and, together with the Gln4 side-chain, and perhaps also the Glu3 side-chain [21], is involved in directing the reactive glutamine to the active site in the substrate-binding cavity. On the basis of indirect evidence provided by 1D proton NMR line broadening and 2D transferred NOESY spectra of N1-a 2 PI (1-15), Marinescu et al [20] suggested an interaction between the C-terminal part of the substrate peptide and FXIIIa. In our study, the existence of a second substratebinding site, important for the transglutaminase reaction, was clearly proven by kinetic experiments with C-terminally truncated/substituted substrates and STD NMR.…”

Section: Discussionmentioning

confidence: 99%

Molecular mechanism of the interaction between activated factor XIII and its glutamine donor peptide substrate

Pénzes¹,

Kövér²,

Fazakas

et al. 2009

Journal of Thrombosis and Haemostasis

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To cite this article: Pé nzes K, Kö vé r KE, Fazakas F, Haramura G, Muszbek L. Molecular mechanism of the interaction between activated factor XIII and its glutamine donor peptide substrate. J Thromb Haemost 2009; 7: 627-33.Summary. Background: Activated factor XIII (FXIII), a dimer of truncated A-subunits (FXIII-A 2 *), is a transglutaminase that crosslinks primary amines to peptide-bound glutamine residues. Because in the few natural substrates of FXIII-A 2 * no consensus sequence could be identified around the reactive glutamine, studying the interaction between individual substrates and FXIII-A 2 * is of primary importance. Most of the a 2 -plasmin inhibitor (a 2 PI) molecules become truncated by a plasma protease, and the truncated isoform (N1-a 2 PI) is an important substrate of FXIII-A 2 *. The crosslinking of N1-a 2 PI to fibrin plays a major role in protecting fibrin from fibrinolysis. Methods: We studied the interaction of FXIII-A 2 * with its dodecapeptide glutamine donor substrate, N1-a 2 PI(1-12), the sequence of which corresponds to the N-terminal sequence of N1-a 2 PI. Kinetic parameters for N1-a 2 PI(1-12) and for its truncated or synthetic mutants were determined by a spectrophotometric assay. The interaction of N1-a 2 PI(1-12) with FXIII-A 2 * was investigated by proton nuclear magnetic resonance (NMR) and saturating transfer difference (STD) NMR. Results and Conclusions: Kinetic experiments with peptides in which the Asn1 residue was either truncated or replaced by alanine and proton NMR analysis of the FXIII-A 2 *-N1-a 2 PI(1-12) complex demonstrated that Asn1 is essential for effective enzyme-substrate interaction. Experiments with C-terminally truncated peptides proved that amino acids 7-12 are essential for the interaction of N1-a 2 PI(1-12) with the enzyme, and suggested the existence of a secondary binding site on FXIII-A 2 *. Hydrophobic residues, particularly Leu10 and the C-terminal Lys12, seemed to be especially important in this respect, and direct interaction between hydrophobic C-terminal residues and FXIII-A 2 * was demonstrated by STD NMR.

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“…In recent years several peptides based either on b-casein and a2-antiplasmin for the Gln-donor substrate requirements (21,22) or on the bovine aA-crystallin for the amine-donor substrate requirements have been investigated as model systems for the transglutaminase activity (23).…”

Section: Design and Modeling Of The Imaging Probementioning

confidence: 99%

Novel MRI and fluorescent probes responsive to the Factor XIII transglutaminase activity

Tei¹,

Mazooz²,

Shellef³

et al. 2010

Contrast Media & Molecular

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Transglutaminases, including factor XIII and tissue transglutaminase, participate in multiple extracellular processes associated with remodeling of the extracellular matrix during wound repair, blood clotting, tumor progression and fibrosis of ischemic injuries. The aim of this work was to evaluate a novel substrate analog for transglutaminase optimized by molecular modeling calculations (DCCP16), which can serve for molecular imaging of transglutaminase activity by magnetic resonance imaging and by near-infrared imaging. Experimental data showed covalent binding of Gd-DCCP16 and DCCP16-IRIS Blue to human clots, to basement membrane components and to casein in purified systems as well as in three-dimensional multicellular spheroids. In vivo, DCCP16 showed enhancement with a prolonged retention in clots and tumors, demonstrating the ability to detect both factor XIII and tissue transglutaminase mediated covalent binding of the contrast material.

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Structural features associated with the binding of glutamine-containing peptides to Factor XIII

Cited by 11 publications

References 35 publications

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Molecular mechanism of the interaction between activated factor XIII and its glutamine donor peptide substrate

Novel MRI and fluorescent probes responsive to the Factor XIII transglutaminase activity

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