PURPOSE: Myocilin is thought to be a stress response protein, but its exact molecular functions have not been established. Studies were conducted to see whether myocilin can act as a general molecular chaperone. METHODS: Myocilin was isolated and purified from porcine trabecular meshwork (TM) cell culture media. Its ability to protect citrate synthase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the restriction endonuclease DrdI from thermal inactivation was evaluated. Light scattering was used to evaluate thermally induced aggregation of citrate synthase. Myocilin induction was assessed after exposure of TM cells to several types of stress treatments. RESULTS: Levels of extracellular myocilin expressed by TM cells were increased in response to mechanical stretch, heat shock, TNFα, or IL-1α. Myocilin protected citrate synthase activity against thermal inactivation for 5 minutes at 55°C in a concentration-dependent manner, with nearly full protection of 1.5 μM citrate synthase in the presence of 650 nM myocilin. Myocilin significantly reduced thermal aggregation of citrate synthase to levels 36% to 44% of control levels. Myocilin also protected GAPDH from thermal inactivation for 10 minutes at 45°C. Myocilin at 18 nM was more effective than 1 μM bovine serum albumin at protecting DrdI from thermal inactivation. CONCLUSIONS: Myocilin is induced in response to several cellular stresses and displays general molecular chaperone activity by protecting DrdI, citrate synthase, and GAPDH from thermal inactivation. Myocilin also suppresses the thermal aggregation of citrate synthase. One function of myocilin may be to serve as a molecular chaperone.
PURPOSE:Myocilin is thought to be a stress response protein, but its exact molecular functions have not been established. Studies were conducted to see whether myocilin can act as a general molecular chaperone. METHODS:Myocilin was isolated and purified from porcine trabecular meshwork (TM) cell culture media. Its ability to protect citrate synthase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the restriction endonuclease DrdI from thermal inactivation was evaluated. Light scattering was used to evaluate thermally induced aggregation of citrate synthase. Myocilin induction was assessed after exposure of TM cells to several types of stress treatments. RESULTS: Levels of extracellular myocilin expressed by TM cells were increased in response to mechanical stretch, heat shock, TNFα, or IL-1α. Myocilin protected citrate synthase activity against thermal inactivation for 5 minutes at 55°C in a concentration-dependent manner, with nearly full protection of 1.5 μM citrate synthase in the presence of 650 nM myocilin. Myocilin significantly reduced thermal aggregation of citrate synthase to levels 36% to 44% of control levels. Myocilin also protected GAPDH from thermal inactivation for 10 minutes at 45°C. Myocilin at 18 nM was more effective than 1 μM bovine serum albumin at protecting DrdI from thermal inactivation. CONCLUSIONS:Myocilin is induced in response to several cellular stresses and displays general molecular chaperone activity by protecting DrdI, citrate synthase, and GAPDH from thermal inactivation. Myocilin also suppresses the thermal aggregation of citrate synthase. One function of myocilin may be to serve as a molecular chaperone.
Authors: Susan D Orwig; Pamela V Chi; Yuhong Du; Shannon E Hill; Marchello A Cavitt; Amrithaa Suntharalingam; Katherine C Turnage; Chad A Dickey; Stefan France; Haian Fu; Raquel L Lieberman Journal: ACS Chem Biol Date: 2013-12-09 Impact factor: 5.100
Authors: Kate E Keller; Sanjoy K Bhattacharya; Theresa Borrás; Thomas M Brunner; Sunee Chansangpetch; Abbott F Clark; W Michael Dismuke; Yiqin Du; Michael H Elliott; C Ross Ethier; Jennifer A Faralli; Thomas F Freddo; Rudolf Fuchshofer; Michael Giovingo; Haiyan Gong; Pedro Gonzalez; Alex Huang; Murray A Johnstone; Paul L Kaufman; Mary J Kelley; Paul A Knepper; Casey C Kopczynski; John G Kuchtey; Rachel W Kuchtey; Markus H Kuehn; Raquel L Lieberman; Shan C Lin; Paloma Liton; Yutao Liu; Elke Lütjen-Drecoll; Weiming Mao; Marisse Masis-Solano; Fiona McDonnell; Colleen M McDowell; Darryl R Overby; Padmanabhan P Pattabiraman; Vijay K Raghunathan; P Vasanth Rao; Douglas J Rhee; Uttio Roy Chowdhury; Paul Russell; John R Samples; Donald Schwartz; Evan B Stubbs; Ernst R Tamm; James C Tan; Carol B Toris; Karen Y Torrejon; Janice A Vranka; Mary K Wirtz; Thomas Yorio; Jie Zhang; Gulab S Zode; Michael P Fautsch; Donna M Peters; Ted S Acott; W Daniel Stamer Journal: Exp Eye Res Date: 2018-03-09 Impact factor: 3.770