ABSTRACT
The phenomenon that SARS coronavirus main protease (SARS M(pro)) dimer is the main functional form has been confirmed by experiment. However, because of the absence of structural information of the monomer, the reasons for this remain unknown. To investigate it, two molecular dynamics (MD) simulations in water for dimer and monomer models have been carried out, using the crystal structure of protomer A of the dimer as the starting structure for the monomer. During the MD simulation of dimer, three interest phenomena of protomer A have been observed: (i) the distance between NE2 of His41 and SG of Cys145 averages 3.72 A, which agrees well with the experimental observations made by X-ray crystallography; (ii) His163 and Glu166 form the "tooth" conformational properties, resulting in the specificity for glutamine at substrate P1 site; and (iii) the substrate-binding pocket formed by loop 140-146 and loop 184-197 is large enough to accommodate the substrate analog. However, during the MD simulation of the monomer complex, the three structural characteristics are all absent, which results directly in the inactivation of the monomer. Throughout the MD simulation of the dimer, the N-terminus of protomer B forms stable hydrogen bonds with Phe140 and Glu166, through which His163, Glu166, and loop 140-146 are kept active form. Furthermore, a water-bridge has been found between the N-terminus of protomer B and Gly170, which stabilizes His172 and avoids it moving toward Tyr161 to disrupt the H-bond between Tyr161 and His163, stabilizing the conformation of His163. The interactions between the N-terminus and another monomer maintain the activity of dimer.
Subject(s)
Computer Simulation , Cysteine Endopeptidases/metabolism , Models, Chemical , Severe acute respiratory syndrome-related coronavirus/enzymology , Viral Proteins/metabolism , Catalytic Domain , Coronavirus 3C Proteases , Cysteine Endopeptidases/chemistry , Dimerization , Hydrogen Bonding , Hydrolysis , Models, Molecular , Protease Inhibitors/pharmacology , Protein Conformation , Protein Interaction Mapping , Structure-Activity Relationship , Substrate Specificity , Viral Proteins/chemistry , WaterABSTRACT
Maternal immunoglobulin G (IgG) was derived from Wistar rats that just delivered the new offsprings. We examined the effect of this maternal IgG on infantile spasms induced by N-methyl-d-aspartate (NMDA) in immature rats. Pup animals were treated subcutaneously with 10 mg/kg/day maternal IgG from day 11 to day 15 after birth followed by a single intraperitoneal dose of NMDA (15 mg/kg). Administration of maternal IgG decreased the severity and increased the number of ACTH immunoreactive cells in the cortex of rats with NMDA-induced spasms. Furthermore, maternal IgG inhibited NMDA-induced intracellular LDH activity in cultured hippocampal neurons in a dose-dependent manner. The results indicate that maternal IgG can attenuate NMDA-induced seizures. In infantile spasms, some factors may during pregnancy negatively affect the transfer of maternal IgG from mother to fetus thereby causing a decrease in the amount of protective maternal IgG.
