Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study.

Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Research Abstract Details 

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Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Abstract Text:

Giulia Isetti,Muriel C Maurer,

In the last stages of coagulation, thrombin helps to activate Factor XIII. The resultant transglutaminase introduces covalent cross-links into fibrin thus promoting clot stability. To better understand the roles of individual thrombin residues in recognition and hydrolysis of the Factor XIII activation peptide, mutations within thrombin's aryl and apolar binding site were explored. The thrombin mutants W215A, E217A, W215A/E217A, L99A, and I174A were examined through HPLC kinetics against the substrates FXIII (28-41) V34 AP and FXIII (28-41) V34L AP. Several mutants responded differently to FXIII (28-41) V34 AP vs the cardioprotective V34L AP. W215 provides an important platform for binding and directing FXIII APs for proper hydrolysis. Loss of this platform leads to decreases in kinetics, particularly to the kcat of FXIII V34L AP. E217 also plays a supporting role, but the E217A mutation is not as detrimental as W215A. W215A/E217A is unfavorable for both activation peptides and its coupling effect has been characterized. This mutant can readily bind the peptides but cannot orient them for effective hydrolysis. Kinetic studies with I174A indicate that this thrombin residue is more crucial for interactions with the larger V34L AP segment. The L99A mutation causes deleterious effects to binding and hydrolysis of both APs. The V34L, however, is able to partially compensate for the loss perhaps by increasing contact within the aryl and apolar sites. Understanding how specific FXIII and thrombin residues participate in binding and control hydrolysis may lead to the design of coagulation enzymes whose degree of activation and optimal target site can be controlled.

Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Publishing Authors By Initials

G Isetti,MC Maurer,

For similar enzymes and coenzymes: enzymes: hydrolases: peptide hydrolases: endopeptidases: serine endopeptidases: thrombin research abstracts see: enzymes and coenzymes: enzymes: hydrolases: peptide hydrolases: endopeptidases: serine endopeptidases: thrombin research

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MEDLINE DATE:

Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Biochemistry

VOLUME: 46

Page Numbers: 2444-52

Journal Abbreviation: Biochemistry

ISSN: 0006-2960

DAY: 8

MONTH: 02

YEAR: 2007

Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 370623

Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Keywords Mesh Terms:

KEYWORDS: Thrombin

MESH TERMS: metabolism

Chemical & Substance for Abstract: Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study. Information

Substance Name: Thrombin

Registry Number: EC 3.4.21.5

Grant and Affiliation Information for Employing mutants to study thrombin residues responsible for factor XIII activation peptide recognition: a kinetic study.

AFFILIATION: Department of Chemistry, University of Louisville, 2320 South Brook Street, Louisville, Kentucky 40292, USA.

Country: United States

AGENCY: United States NHLBI

GRANT: R01 HL68440

ACRONYM: HL

MEDLINETA: Biochemistry

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