Assembly and lysine binding of recombinant lipoprotein(a)

Ernst, Angelika; Brunner, Christoph; Schramm, Axel; Helmhold, M.; Armstrong, Victor W.; ̈ller, H.-J. Mu

doi:10.1016/0021-9150(94)94385-0

Cited by 2 publications

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“…These data indicate that the main functional role of KIV-2 cannot be related to a classical lysine binding mechanism mediated by an optimized LBS and is in line with previous sequence-based hypothesis and experimental findings (28). In fact, structural differences generate an "affinity gradient" for EACA going from the high-affinity C-terminal KIV-10 to the low affinity Nterminal KIV-2 of apo(a).…”

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confidence: 90%

“…In fact, a dominant role for KIV-10 in the lysine-binding function of apo(a) was first demonstrated by the loss of binding observed in a KIV-10 W70R mutant of Rhesus monkey (27). Further lysine binding capacity was later found localised within the KIV-5 to KIV-9 domains, while the KIV-1 to KIV-4 domains did not show any significant lysine binding capacity (28). Within the KIV-5 to KIV-9 range, domains KIV-6 and KIV-7 have a crucial role in Lp(a) assembly (29)(30)(31)(32) and their LBS shows moderate affinity for the lysine analogue -aminocaproic acid (EACA, 0.2-0.3 mM, see Table 1).…”

Section: Introductionmentioning

confidence: 95%

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High resolution structure of human apolipoprotein (a) kringle IV type 2: beyond the lysine binding site

Santonastaso

Maggi

Jonge

et al. 2020

Journal of Lipid Research

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Lipoprotein (a) [Lp(a)] is characterized by an LDL-like composition in terms of lipids and apoB100, and by one copy of a unique glycoprotein, apolipoprotein (a) [apo(a)]. The apo(a) structure is mainly based on the repetition of tandem kringle domains with high homology to plasminogen kringles 4 and 5. Among them, kringle IV type 2 (KIV-2) is present in a highly variable number of genetically encoded repeats, whose length is inversely related to Lp(a) plasma concentration and cardiovascular risk. Despite it being the major component of apo(a), the actual function of KIV-2 is still unclear. Here, we describe the first high-resolution crystallographic structure of this domain. It shows a general fold very similar to other KIV domains with high and intermediate affinity for the lysine analogue ε-aminocaproic acid (EACA). Interestingly, KIV-2 presents a Lysine Binding Site (LBS) with a unique shape and charge distribution. KIV-2 affinity for predicted small molecule binders was found to be negligible in Surface Plasmon Resonance (SPR) experiments, and with the LBS being nonfunctional, we propose to rename it “pseudo-LBS” (pLBS). Further investigation of the protein by computational small-molecule docking allowed us to identify a possible heparin-binding site away from the LBS, which was confirmed by specific reverse charge mutations abolishing heparin binding. This study opens new possibilities to define the pathogenesis of Lp(a) related diseases and to facilitate the design of specific therapeutic drugs. (226 words)

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confidence: 90%

Section: Introductionmentioning

confidence: 95%

High resolution structure of human apolipoprotein (a) kringle IV type 2: beyond the lysine binding site

Santonastaso

Maggi

Jonge

et al. 2020

Journal of Lipid Research

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“…These developments have broadened the search for possible pathological roles for Lp [a] to include thrombosis in addition to atherosclerosis (7,8). Kringle structures possess specific ligand-binding properties, with documented binding sites for l -lysine and related amino acid analogs (9,10). Ten distinctive kringle IV types have been found in apo[a]; certain types have been identified in the specific noncovalent interaction with apoB-100, which is the first step in Lp [a] particle assembly.…”

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confidence: 99%

Isolation, quantitation, and characterization of a stable complex formed by Lp[a] binding to triglyceride-rich lipoproteins

Gaubatz

Hoogeveen

Hoffman

et al. 2001

Journal of Lipid Research

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Lipoprotein [a] (Lp[a]) is a cholesterol-rich lipoprotein resembling LDL to which a large polymorphic glycoprotein, apolipoprotein [a] (apo[a]), is covalently coupled. Lp[a] usually exists as a free-standing particle in normolipidemic subjects; however, it can associate noncovalently with triglyceride-rich lipoproteins in hypertriglyceridemic (HTG) subjects. In this study, 10-78% of the Lp[a] present in five HTG subjects was found in the triglyceride-rich lipoprotein (TRL) fraction. The Lp[a]-TRL complex was resistant to dissociation by ultracentrifugation (UCF) alone, but was quantitatively dissociated by UCF in the presence of 100 mM proline. Of this dissociated Lp[a], 70-88% was in the form of a lipoprotein resembling conventional Lp[a]. Incubation of Lp[a]-depleted TRL with native Lp[a] resulted in a reconstituted Lp[a]-TRL complex that closely resembled the native isolates in all examined properties. Complex formation was inhibited by several compounds in the order proline Ͼ tranexamate Ͼ -aminocaproate ϾϾ arginine Ͼ lysine. Neither plasminogen nor LDL inhibited binding of Lp[a] to TRL. We observed the preferential binding of Lp[a] containing higher apparent molecular weight apo[a] polymorphs to TRL both in native and reconstituted Lp[a]-TRL complexes. A disproportionate amount of Lp[a] was bound to the larger TRL particles. Although most apo[a] bound to TRL was in the form of conventional Lp[a] particles, lipidfree recombinant apo[a] was observed to bind TRL. These results provide unequivocal evidence of the existence of an Lp[a]-TRL complex under pathophysiologic conditions. The metabolic fate of the Lp[a]-TRL complex, which is more abundant in hypertriglyceridemia, may be different from that of conventional Lp[a], and may contribute uniquely to the progression or severity of cardiovascular disease.

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Assembly and lysine binding of recombinant lipoprotein(a)

Cited by 2 publications

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High resolution structure of human apolipoprotein (a) kringle IV type 2: beyond the lysine binding site

High resolution structure of human apolipoprotein (a) kringle IV type 2: beyond the lysine binding site

Isolation, quantitation, and characterization of a stable complex formed by Lp[a] binding to triglyceride-rich lipoproteins

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