Mahmood Akhtar, Engle Michael J, Alpers David H
Division of Gastroenterology, Washington University School of Medicine, St Louis, MO 63110, USA.
J Physiol. 2002 Jul 1;542(Pt 1):237-44. doi: 10.1113/jphysiol.2002.017087.
Surfactant-like particles (SLP) are secreted from enterocytes basolaterally into the lamina propria, and reach the apical surface through the intercellular tight junctions. Interactions of SLP with apical and basolateral membranes and with extracellular matrix proteins were measured using a solid-phase binding assay and gel overlays. Small-intestinal SLP bound to basolateral membranes much more than to apical membranes, and more tightly to fibronectin than to laminin (affinity constant K(a) = 1.23 x 10(-2) microg vs. 0.67 x 10(-2) microg; maximal number of binding sites 4.1 microg x ml(-1) vs. 0.32 microg x ml(-1)), but did not bind to collagen types I or IV. Small-intestinal SLP bound fibronectin more than colonic or gastric SLP. Binding to fibronectin was inhibited only partially by RGD peptide and gelatin, but not by heparin. An antibody against alpha(v) integrin also identified the fibronectin-binding component in SLP at approximately 220 kDa, which is the expected size for integrin heterodimers. SLP binding to apical microvillous membranes was weaker and was inhibited by heparin. SLP bound more strongly to heparin itself, and this binding was inhibited by glucuronic acid and chondroitin sulfate. These data are consistent with the hypothesis that the time spent by secreted SLP in the lamina propria is prolonged by strong interactions with proteins in the basolateral membranes, and in the intestinal lumen by weaker interactions with apical membrane components, including heparin. These interactions may allow SLP the time to exert their functions in each tissue compartment.
表面活性剂样颗粒(SLP)从肠上皮细胞基底外侧分泌到固有层,并通过细胞间紧密连接到达顶端表面。使用固相结合测定法和凝胶覆盖法测量了SLP与顶端和基底外侧膜以及细胞外基质蛋白的相互作用。小肠SLP与基底外侧膜的结合远多于与顶端膜的结合,与纤连蛋白的结合比与层粘连蛋白的结合更紧密(亲和常数Ka = 1.23×10⁻²微克对0.67×10⁻²微克;最大结合位点数4.1微克×毫升⁻¹对0.32微克×毫升⁻¹),但不与I型或IV型胶原结合。小肠SLP比结肠或胃SLP与纤连蛋白的结合更多。与纤连蛋白的结合仅部分受到RGD肽和明胶的抑制,但不受肝素的抑制。一种针对α(v)整合素的抗体也在约220 kDa处鉴定出SLP中的纤连蛋白结合成分,这是整合素异二聚体的预期大小。SLP与顶端微绒毛膜的结合较弱且受到肝素的抑制。SLP与肝素本身的结合更强,并且这种结合受到葡萄糖醛酸和硫酸软骨素的抑制。这些数据与以下假设一致,即分泌的SLP在固有层中与基底外侧膜中的蛋白质强烈相互作用从而延长其停留时间,而在肠腔中则通过与顶端膜成分(包括肝素)的较弱相互作用延长停留时间。这些相互作用可能使SLP有时间在每个组织隔室中发挥其功能。