Structural and Biophysical Aspects of Lactoferrin and Its
Interaction with Plasminogen
Patricia Hrasnova, Klaudia Meskova, Rostislav Skrabana
Institute of Neuroimmunology, Slovak Academy of Sciences
Recently it was shown that lactoferrin (Lf),
immunomodulatory protein of transferrin family of proteins interacts with
plasminogen (Plg), protein of the fibrinolytic cascade
. The most probable mode of interaction is the contact of the N-terminal
domain of Lf (corresponding to its peptide lactoferricin) with a mini-Plg
region consisting of kringle 5 and a proteolytic domain.
Our aim is to further characterise the properties of Lf, comparing its recombinant and natural form.
Unlike the native Lf, commercially available recombinant Lf produced in
transgenic rice bears different type of glycosylation. Therefore, we aimed to
prepare a recombinant Lf with human-like glycosylation
by its expression in mammalian CHO cell line.
Depending on the environment, Lf may oligomerise, losing
its bactericidal and fungicidal immunomodulatory characteristics [2, 3]. Some
properties of Lf can be changed in the presence of divalent ions (especially calcium ions ) inducing aggregation into higher
oligomeric states. We used dynamic light scattering analysis to characterise Lf
Previous results indicated that the functional part of Lf
interaction with Plg consists of the peptide
lactoferricin . Our next goal was to optimise its production by pepsin
digestion and its subsequent isolation. To determine the exact interaction of
prepared lactoferricin with Plg, affinity determination by surface plasmon
resonance will be used. Additionally to these
experimental approaches, in silico molecular docking characterising the interaction sites will be applied.
Figure 1: Plasminogen (Plg)
activation to plasmin (Plm) by tissue-type plasminogen activator (tPA) and
urokinase plasminogen activator (uPA) (modified from ) and its regulation by
melanotransferrin (Mtf) and lactoferrin (Lf ).
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