Identification of amino acid residues that are crucial for FXIII-A intersubunit interactions and stability.

Li, Bojun; Kohler, Hans P.; Schroeder, Verena (2020). Identification of amino acid residues that are crucial for FXIII-A intersubunit interactions and stability. Blood, 135(2), pp. 145-152. American Society of Hematology 10.1182/blood.2019002127

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Coagulation factor XIII (FXIII) is the main stabilizer of the fibrin clot. It circulates in plasma as a tetramer of two A and two B subunits. Under physiological conditions, FXIII-A exists as a dimer (FXIII-A2). The interactions between the FXIII-A subunits that stabilize the FXIII-A2 dimer are not fully understood. Therefore, we designed a systematic approach to identify amino acid residues crucial for the expression and stability of FXIII-A2. Based on the available FXIII-A2 crystal structure, we identified 12 amino acid residues forming intersubunit salt bridges and 21 amino acid residues forming hydrogen bonds between the two A-subunits. We chose ten amino acid residues that form five particularly strong interactions, performed site-directed mutagenesis and expressed the mutants in CHO cells. Disruption of these interactions by single mutation of Lys257, Lys113, Asp343, Glu401, or Asp404 abolished the expression of properly folded, soluble and functional FXIII-A in CHO cells. On the contrary, mutation of Glu111, Arg100, or Asn112 had no significant effect on FXIII-A expression. Our results suggest that four intersubunit interactions (Arg11-Asp343, Lys113-Asp367, Lys257-Glu401 and Arg260-Asp404) are essential for the stability of FXIII-A2. Our findings are supported by reported mutations at Lys257, Arg260, and Asp404 found in patients with congenital FXIII-A deficiency.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Pavillon 52 > Forschungsgruppe Experimentelle Hämostase
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)

UniBE Contributor:

Li, Bojun; Kohler, Hans-Peter and Schröder, Verena

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0006-4971

Publisher:

American Society of Hematology

Language:

English

Submitter:

Marla Rittiner

Date Deposited:

27 Dec 2019 13:52

Last Modified:

23 Oct 2020 02:30

Publisher DOI:

10.1182/blood.2019002127

PubMed ID:

31697820

BORIS DOI:

10.7892/boris.136261

URI:

https://boris.unibe.ch/id/eprint/136261

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