Journal of Biological Chemistry · 1992 · 155 citations · 43 references
Life SciencesBiochemistryMolecular BiologyEnzymologyLipid BiochemistryProtein BiochemistryLipid-protein InteractionsMilk EnzymesMilk LipidsProtein BindingProtein-ligand BindingProtein-lipid InteractionsLipolytic ActivityLipoprotein Lipase FunctionLipoprotein Lipase-mediated LipolysisLipoprotein ReceptorsMilk LipoproteinsPolyclonal Antibody RecognitionReceptorsTriglyceride-rich Lipoproteins
Lipoprotein lipase (LPL), the major lipolytic enzyme involved in the conversion of triglyceride-rich lipoproteins to remnants, was found to compete with binding of activated alpha 2-macroglobulin (alpha 2M*) to the low density lipoprotein receptor-related protein (LRP)/alpha 2-macroglobulin receptor. Bovine milk LPL displaced both 125I-labeled alpha 2M* and 39-kDa alpha 2M receptor-associated protein (RAP) from the surface of cultured mutant fibroblasts lacking LDL receptors with apparent KI values at 4 degrees C of 6.8 and 30 nM, respectively. Furthermore, LPL inhibited the cellular degradation of 125I-alpha 2M* at 37 degrees C. Because both alpha 2M* and RAP interact with LRP, these data suggest that LPL binds specifically to this receptor. This was further supported by observing that an immunoaffinity-isolated polyclonal antibody against LRP blocked cellular degradation of 125I-LPL in a dose-dependent manner. In addition, 125I-LPL bound to highly purified LRP in a solid-phase assay with a KD of 18 nM, and this binding could be partially displaced with alpha 2M* (KI = 7 nM) and RAP (KI = 3 nM). Taken together, these data establish that LPL binds with high affinity to LRP and undergoes LRP-mediated cellular uptake. The implication of these findings for lipoprotein catabolism in vivo may be important if LRP binding is preserved when LPL is attached to lipoproteins. If so, LPL might facilitate LRP-mediated clearance of lipoproteins.
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Joachim Herz, Ute Hamann, Sissel Rogne et al. · The EMBO Journal · 1988 · 958 citations · Full text
A new solid-state reagent to iodinate proteins
Mary Ann K. Markwell · Analytical Biochemistry · 1982 · 877 citations