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Louis, Missouri 1 To whom correspondence should be addressed: Louis University School of Medicine, St. This article has been cited by other articles in PMC. Abstract A flexible linker Lnk2 composed of 26 amino acids connects kringle-1 to kringle-2 in the coagulation factor prothrombin. Recent studies point to Lnk2 as a key determinant of the structure and function of this zymogen. Using a combination of mutagenesis, structural biology, and single molecule spectroscopy, we show how Lnk2 influences activation and conformational plasticity of prothrombin. Scrambling the sequence of Lnk2 is inconsequential on activation, and so is extension by as many as 22 residues. On the other hand, below a critical length of 15 residues, the rate of prothrombin activation increases fold in the absence of cofactor Va and decreases 3-fold in the presence of cofactor. Furthermore, activation by prothrombinase takes place without preference along the prethrombin-2 cleavage at Arg first or meizothrombin cleavage at Arg first pathways. Notably, these transitions in the rate and pathway of activation require the presence of phospholipids, pointing to an important physiological role for Lnk2 when prothrombin is anchored to the membrane.

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