Probing thrombin's ability to accommodate a V34F substitution within the factor XIII activation peptide segment (28–41)*

Isetti, Giulia; Maurer, Muriel C.

doi:10.1111/j.1399-3011.2004.00132.x

Cited by 8 publications

(27 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Solution NMR studies have further supported these observations [21, 35, 36]. A hallmark P 4 (L34) to P 2 (P36) through-space NOE interaction likely contributes to enhanced binding of the FXIII V34L AP segment on to the thrombin active surface.…”

Section: Discussionmentioning

confidence: 87%

“…The two prolines at the P 4 and P 2 positions of PAR4 supply critical anchor points on to the thrombin surface and enhance kinetic properties. Earlier kinetic and NMR studies with FXIII (28–41) V34F and thrombin revealed that the aromatic F is the only other residue thus far that can achieve the stabilizing P 4 -P 2 through space interaction observed with L34 [35]. …”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Design of Factor XIII V34X activation peptides to control ability to interact with thrombin mutants

Jadhav

Lucas

Goldsberry

et al. 2011

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

View full text Add to dashboard Cite

Thrombin helps to activate Factor XIII (FXIII) by hydrolyzing the R37-G38 peptide bond. The resultant transglutaminase introduces cross-links into the fibrin clot. With the development of therapeutic coagulation factors, there is a need to better understand interactions involving FXIII. Such knowledge will help predict ability to activate FXIII and thus ability to promote/hinder the generation of transglutaminase activity. Kinetic parameters have been determined for a series of thrombin species hydrolyzing the FXIII (28′41) V34X activation peptides (V34, V34L, V34F, and V34P). The V34P substitution introduces PAR4 character into the FXIII, and the V34F exhibits important similarities to the cardioprotective V34L. FXIII activation peptides containing V34, V34L, or V34P could each be accommodated by alanine mutants of thrombin lacking either the W60d or Y60a residue in the 60-insertion loop. By contrast, FXIII V34F AP could be cleaved by thrombin W60dA but not by Y60aA. FXIII V34P is highly reliant on the thrombin W215 platform for its strong substrate properties whereas FXIII V34F AP becomes the first segment that can maintain its Km upon loss of the critical thrombin W215 residue. Interestingly, FXIII V34F AP could also be readily accommodated by thrombin L99A and E217A. Hydrolysis of FXIII V34F AP by thrombin W217A/E217A (WE) was similar to that of FXIII V34L AP whereas WE could not effectively cleave FXIII V34P AP. FXIII V34F and V34P AP show promise for designing FXIII activation systems that are either tolerant of or greatly hindered by the presence of anticoagulant thrombins.

show abstract

Section: Discussionmentioning

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Design of Factor XIII V34X activation peptides to control ability to interact with thrombin mutants

Jadhav

Lucas

Goldsberry

et al. 2011

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

View full text Add to dashboard Cite

show abstract

“…This structural feature has been documented by both X-ray (33) and solution NMR methods (35, 36). So far a helical turn conformation has not been observed in solution (11, 15) with FXIII AP (28–37) V34, V34L, V34F, V29F, or the doubly substituted V29F, V34L. If such a conformation does exist, it must be fleeting in nature.…”

Section: Discussionmentioning

confidence: 99%

Effects of Introducing Fibrinogen Aα Character into the Factor XIII Activation Peptide Segment

et al. 2010

View full text Add to dashboard Cite

The formation of a blood clot involves the interplay of thrombin, fibrinogen, and Factor XIII. Thrombin cleaves fibrinopeptides A and B from the N-termini of the fibrinogen Aα and Bβ chains. Fibrin monomers are generated that then polymerize into a noncovalently associated network. By hydrolyzing the Factor XIII activation peptide segment at the R37-G38 peptide bond, thrombin assists in activating the transglutaminase FXIIIa that incorporates cross links into the fibrin clot. In the current research, the kinetic effects of introducing fibrinogen Aα character into the FXIII AP segment were examined. About 25% of fibrinogen Aα is phosphorylated at Ser 3 producing a segment with improved binding to thrombin. FXIII AP (22AEDDL26) has sequence properties in common with Fbg Aα (1ADSpGE5). Kinetic benefits to FXIII AP cleavage were explored by extending FXIII AP (28–41) to FXIII AP (22–41) and examining peptides with D24, D24S, D24Sp, and D24Sp P27G. These modifications did not provide the same kinetic advantages as observed with Fbg Aα (1–20) S3p. Such results further emphasize that FXIII AP derives most of its substrate specificity from the P9-P1 segment. To enhance the kinetic properties of FXIII AP (28–41), substitutions were introduced at the P9, P4, and P3 positions. Studies reveal that FXIII AP (28–41) V29F, V34G, V35G exhibits kinetic improvements that are comparable to FXIII AP V29F, V34L and approach those of Fbg Aα (7–20). Selective changes to the FXIII AP segment sequence may be used to design FXIII species that can be activated more or less readily.

show abstract

“…The FXIII P34 variant allowed platelet PAR4 character to be introduced into the FXIII AP sequence (Table 1) [47]. Earlier NMR studies had revealed that the FXIII F34 provided the same beneficial P 4 -P 2 interactions with the thrombin surface as L34 [39]. Importantly, the FXIII F34 AP was demonstrated to be the first FXIII-based substrate that exhibited little change in K m upon loss of thrombin residue W215 [40].…”

Section: Introductionmentioning

confidence: 99%

Screening cleavage of Factor XIII V34X Activation Peptides by thrombin mutants: A strategy for controlling fibrin architecture

Jadhav

Goldsberry

Zink

et al. 2017

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

View full text Add to dashboard Cite

In blood coagulation, thrombin converts fibrinogen into fibrin monomers that polymerize into a clot network. Thrombin also activates Factor XIII by cleaving the R37-G38 peptide bond of the Activation Peptide (AP) segment. The resultant transglutaminase introduces covalent crosslinks into the fibrin clot. A strategy to modify clot architecture would be to design FXIII AP sequences that are easier or more difficult to be thrombin-cleaved thus controlling initiation of crosslinking. To aid in this design process, FXIII V34X (28–41) activation peptides were kinetically ranked for cleavage by wild-type thrombin and several anticoagulant mutants. Thrombin-catalyzed hydrolysis of aromatic FXIII F34, W34, and Y34 APs was compared with V34 and L34. Cardioprotective FXIII L34 remained the variant most readily cleaved by wild-type thrombin. The potent anticoagulant thrombins W215A and W215A/E217A (missing a key substrate platform for binding fibrinogen) were best able to hydrolyze FXIII F34 and W34 APs. Thrombin I174A and L99A could effectively accommodate FXIII W34 and Y34 APs yielding kinetic parameters comparable to FXIII AP L34 with wild-type thrombin. None of the aromatic FXIII V34X APs could be hydrolyzed by thrombin Y60aA. FXIII F34 and W34 are promising candidates for FXIII – anticoagulant thrombin systems that could permit FXIII-catalyzed crosslinking in the presence of reduced fibrin formation. By contrast, FXIII Y34 with thrombin (Y60aA or W215A/E217A) could help assure that both fibrin clot formation and protein crosslinking are hindered. Regulating the activation of FXIII is predicted to be a strategy for helping to control fibrin clot architecture and its neighboring environments.

show abstract

Probing thrombin's ability to accommodate a V34F substitution within the factor XIII activation peptide segment (28–41)*

Cited by 8 publications

References 41 publications

Design of Factor XIII V34X activation peptides to control ability to interact with thrombin mutants

Design of Factor XIII V34X activation peptides to control ability to interact with thrombin mutants

Effects of Introducing Fibrinogen Aα Character into the Factor XIII Activation Peptide Segment

Screening cleavage of Factor XIII V34X Activation Peptides by thrombin mutants: A strategy for controlling fibrin architecture

Contact Info

Product

Resources

About