Characterizing the specificity of activated Factor XIII for glutamine-containing substrate peptides

Cleary, David B.; Maurer, Muriel C.

doi:10.1016/j.bbapap.2006.05.003

Cited by 33 publications

(54 citation statements)

References 40 publications

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“…Sequences based on a 2 AP (N1, 1-15) have been used to characterize individual glutamines, evaluate roles of surrounding residues, and assess effects of peptide length. [53][54][55] By contrast, similar sized peptides based on the fibrinogen Aa and g chains make poor Q-substrates for FXIIIa. 56,57 These observations suggest that fibrinogen chains require a larger, more protein-like environment to promote substrate specificity.…”

Section: Discussionmentioning

confidence: 99%

Ranking reactive glutamines in the fibrinogen αC region that are targeted by blood coagulant factor XIII

Mouapi

Bell

Smith

et al. 2016

Blood

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Key Points• FXIIIa exhibits a preference for Q237 in crosslinking reactions within fibrinogen aC (233-425) followed by Q328 and Q366.• None of the reactive glutamines in aC 233-425 (Q237, Q328, and Q366) are required to react first before the others can crosslink.Factor XIIIa (FXIIIa) introduces covalent g-glutamyl-«-lysyl crosslinks into the blood clot network. These crosslinks involve both the g and a chains of fibrin. The C-terminal portion of the fibrin a chain extends into the aC region (210-610). Crosslinks within this region help generate a stiffer clot, which is more resistant to fibrinolysis. Fibrinogen aC (233-425) contains a binding site for FXIIIa and three glutamines Q237, Q328, and Q366 that each participate in physiological crosslinking reactions. Although these glutamines were previously identified, their reactivities toward FXIIIa have not been ranked. Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and nuclear magnetic resonance (NMR) methods were thus used to directly characterize these three glutamines and probe for sources of FXIIIa substrate specificity. Glycine ethyl ester (GEE) and ammonium chloride served as replacements for lysine. Mass spectrometry and 2D heteronuclear single quantum coherence NMR revealed that Q237 is rapidly crosslinked first by FXIIIa followed by Q366 and Q328. Both N-GEE could be crosslinked to the three glutamines in aC (233-425) with a similar order of reactivity as observed with the MALDI-TOF mass spectrometry assay. NMR studies using the single aC mutants Q237N, Q328N, and Q366N demonstrated that no glutamine is dependent on another to react first in the series. Moreover, the remaining two glutamines of each mutant were both still reactive. Further characterization of Q237, Q328, and Q366 is important because they are located in a fibrinogen region susceptible to physiological truncations and mutation. The current results suggest that these glutamines play distinct roles in fibrin crosslinking and clot architecture. (Blood. 2016;127(18):2241-2248

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Section: Discussionmentioning

confidence: 99%

Ranking reactive glutamines in the fibrinogen αC region that are targeted by blood coagulant factor XIII

Mouapi

Bell

Smith

et al. 2016

Blood

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“…They showed that both Asn1 in Asn-a2AP (corresponding to Asn13 in Met-a2AP) and amino acids 7 through 12 in Asn-a2AP (corresponding to amino acids 19-24 in Met-a2AP) as a secondary binding site are essential for an effective enzyme-substrate interaction. 52,53 Lee et al 48 also purified and identified the plasma proteinase that is responsible for the in vitro conversion of Met-a2AP to Asn-a2AP and named it antiplasmin-cleaving enzyme. The enzyme appeared to be a soluble, circulating derivative of fibroblast activation protein (FAP) that lacked the cytoplasmic tail and transmembrane part of FAP.…”

Section: N-terminal Variationmentioning

confidence: 99%

Natural heterogeneity of α2-antiplasmin: functional and clinical consequences

Abdul

Leebeek

Rijken

et al. 2016

Blood

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Human a2-antiplasmin (a2AP, also called a2-plasmin inhibitor) is the main physiological inhibitor of the fibrinolytic enzyme plasmin. a2AP inhibits plasmin on the fibrin clot or in the circulation by forming plasmin-antiplasmin complexes. Severely reduced a2AP levels in hereditary a2AP deficiency may lead to bleeding symptoms, whereas increased a2AP levels have been associated with increased thrombotic risk. a2AP is a very heterogeneous protein. In the circulation, a2AP undergoes both amino terminal (N-terminal) and carboxyl terminal (C-terminal) proteolytic modifications that significantly modify its activities. About 70% of a2AP is cleaved at the N terminus by antiplasmin-cleaving enzyme (or soluble fibroblast activation protein), resulting in a 12-amino-acid residue shorter form. The glutamine residue that serves as a substrate for activated factor XIII becomes more efficient after removal of the N terminus, leading to faster crosslinking of a2AP to fibrin and consequently prolonged clot lysis. In approximately 35% of circulating a2AP, the C terminus is absent. This C terminus contains the binding site for plasmin(ogen), the key component necessary for the rapid and efficient inhibitory mechanism of a2AP. Without its C terminus, a2AP can no longer bind to the lysine binding sites of plasmin(ogen) and is only a kinetically slow plasmin inhibitor. Thus, proteolytic modifications of the N and C termini of a2AP constitute major regulatory mechanisms for the inhibitory function of the protein and may therefore have clinical consequences. This review presents recent findings regarding the main aspects of the natural heterogeneity of a2AP with particular focus on the functional and possible clinical implications. (Blood. 2016; 127(5):538-545) Introduction a2-antiplasmin (a2AP, also called a2-plasmin inhibitor) is a key player in the fibrinolytic system (Figure 1). The fibrinolytic system is crucial for dissolving fibrin clots, facilitating tissue repair, and preventing clots from occluding vessels.1 Recent clinical studies have shown that reduced fibrinolysis (eg, due to an increase in a2AP level) is associated with an increase in both venous and arterial thrombotic risk. 2In contrast, an increase in fibrinolysis due to a2AP deficiency is associated with hemophilia-like bleeding symptoms, which typically occur after initial hemostasis as a result of the premature dissolution of fibrin. The phenotype of a2AP deficiency is heterogeneous. Complete congenital a2AP deficiency leads to severe bleeding with hemophilialike bleeding symptoms such as joint bleeding, whereas heterozygous a2AP-deficient patients typically have mild or no bleeding symptoms. 3,4 In addition, acquired a2AP deficiency may also occur in liver disease 5 and amyloidosis 6 or during fibrinolytic therapy. 7The main enzyme of the fibrinolytic system is the serine protease plasmin, which is predominantly responsible for the degradation of fibrin into rapidly cleared soluble fibrin degradation products (Figure 1). The inactive proenzyme plasminogen ca...

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“…The unlabeled control showed no peaks, while the FXIIIa-reacted sample showed the two sharp 15 N-amide proton peaks as anticipated. Similarly, α 2 AP(Q4P) (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15) showed the sharp twin peaks in the spectrum of the FXIII-reacted sample, but the unlabeled control spectrum was not clear like that of K9 (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). In the nonenzymatically reacted sample, there appeared a weak, broad peak at approximately 7 ppm, which was an unexpected result.…”

Section: Fxiiia Interactions With Peptide Substrate Modelsmentioning

confidence: 99%

“…In contrast, K9 (1-10) has its reactive glutamine flanked by glycine and serine residues, both of which only have sp 3 hybridized side chain carbons. This environment appears to not favor the nonenzymatic isotopic exchange, allowing for greater FXIIIa activity.…”

Section: Fxiiia Interactions With αC(233-425)mentioning

confidence: 99%

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Examining the substrate specificity of factor XIIIa towards peptide substrate modelsaC fibrin domains.

Bell

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Characterizing the specificity of activated Factor XIII for glutamine-containing substrate peptides

Cited by 33 publications

References 40 publications

Ranking reactive glutamines in the fibrinogen αC region that are targeted by blood coagulant factor XIII

Ranking reactive glutamines in the fibrinogen αC region that are targeted by blood coagulant factor XIII

Natural heterogeneity of α2-antiplasmin: functional and clinical consequences

Examining the substrate specificity of factor XIIIa towards peptide substrate modelsaC fibrin domains.

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