A tandem repeat of a fragment of Listeria monocytogenes internalin B protein induces cell survival and proliferation

作者：Mungunsukh Ognoon; Lee Young H; Marquez Ana P; Cecchi Fabiola; Bottaro Donald P; Day Regina M^*

来源：American Journal of Physiology - Lung Cellular and Molecular Physiology, 2010, 299(6): L905-L914.

DOI：10.1152/ajplung.00094.2010

• apoptosis
• angiotensin II
• Akt
• ERK1/2

Mungunsukh O, Lee YH, Marquez AP, Cecchi F, Bottaro DP, Day RM. A tandem repeat of a fragment of Listeria monocytogenes internalin B protein induces cell survival and proliferation. Am J Physiol Lung Cell Mol Physiol 299: L905-L914, 2010. First published October 1, 2010; doi:10.1152/ajplung.00094.2010.-Hepatocyte growth factor (HGF) is critical for tissue homeostasis and repair in many organs including the lung, heart, kidney, liver, nervous system, and skin. HGF is a heterodimeric protein containing 20 disulfide bonds distributed among an amino-terminal hairpin, four kringle domains, and a serine protease-like domain. Due to its complex structure, recombinant production of HGF in prokaryotes requires denaturation and refolding, processes that are impractical for large-scale manufacture. Thus, pharmaceutical quantities of HGF are not available despite its potential applications. A fragment of the Listeria monocytogenes internalin B protein from amino acids 36-321 (InlB(36-321)) was demonstrated to bind to and partially activate the HGF receptor Met. InlB(36-321) has a stable beta-sheet structure and is easily produced in its native conformation by Escherichia coli. We cloned InlB(36-321) (1xInlB(36-321)) and engineered a head-to-tail repeat of InlB(36-321) with a linker peptide (2xInlB(36-321)); 1xInlB(36-321) and 2xInlB(36-321) were purified from E. coli. Both 1xand 2xInlB(36-321) activated the Met tyrosine kinase. We subsequently compared signal transduction of the two proteins in primary lung endothelial cells. 2xInlB(36-321) activated ERK1/2, STAT3, and phosphatidylinositol 3-kinase/Akt pathways, whereas 1xInlB(36-321) activated only STAT3 and ERK1/2. The 2xInlB(36-321) promoted improved motility compared with 1xInlB(36-321) and additionally stimulated proliferation equivalent to full-length HGF. Both the 1xand 2xInlB(36-321) prevented apoptosis by the profibrotic peptide angiotensin II in cell culture and ex vivo lung slice cultures. The ease of large-scale production and capacity of 2 x InlB(36-321) to mimic HGF make it a potential candidate as a pharmaceutical agent for tissue repair.

• 出版日期2010-12