title: A nuclear ligand MRG15 involved in the proapoptotic activity of medicinal fungal galectin AAL (Agrocybe aegerita lectin).
abstract: BACKGROUND
  We have previously reported a novel fungal galectin Agrocybe aegerita lectin (AAL) with apoptosis-induced activity and nuclear migration activity. The importance of nuclear localization for AAL's apoptosis-induced activity has been established by mutant study. However, the mechanism remains unclear.
  
  
  METHODS
  We further investigated the mechanism using a previously reported carbohydrate recognition domain (CRD) mutant protein H59Q, which retained its nuclear localization activity but lost most of its apoptotic activity. The cell membrane-binding ability of recombinant AAL (rAAL) and H59Q was analyzed by FACS, and their cellular partners were identified by affinity chromatography and mass spectroscopy. Furthermore, the interaction of AAL and ligand was proved by mammalian two-hybrid and pull down assays. A knockdown assay was used to confirm the role of the ligand.
  
  
  RESULTS
  The apoptotic activity of AAL could be blocked by lactose. Mutant H59Q retained comparable cell membrane-binding ability to rAAL. Four cellular binding partners of AAL in HeLa cells were identified: glucose-regulated protein 78 (GRP78); mortality factor 4-like protein 1 (MRG15); elongation factor 2 (EEF2); and heat shock protein 70 (Hsp70). CRD region of AAL was required for the interaction between AAL/mutant AAL and MRG15. MRG15 knockdown increased the cells' resistance to AAL treatment.
  
  
  CONCLUSIONS
  MRG15 was a nuclear ligand for AAL in HeLa cells. These data implied the existence of a novel nuclear pathway for the antitumor activity of fungal galectin AAL.
  
  
  CONCLUSIONS
  These findings provide a novel explanation of AAL bioactivity and contribute to the understanding of mushroom lectins' antitumor activity.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, People's Republic of China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liang
authors.forename: Jia Cheng
authors.initials: JC
authors.lastname: Lin
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.forename: Yi Jie
authors.initials: YJ
authors.lastname: Chen
authors.forename: Hong Hong
authors.initials: HH
authors.lastname: Liu
authors.forename: Rong
authors.initials: R
authors.lastname: Luan
authors.forename: Shuai
authors.initials: S
authors.lastname: Jiang
authors.forename: Tao
authors.initials: T
authors.lastname: Che
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhao
authors.forename: De Feng
authors.initials: de F
authors.lastname: Li
authors.forename: Da Cheng
authors.initials: da C
authors.lastname: Wang
authors.forename: Lin
authors.initials: L
authors.lastname: Guo
authors.forename: Hui
authors.initials: H
authors.lastname: Sun
mesh_terms: D055362:Agrocybe; D019943:Amino Acid Substitution; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D002846:Chromatography, Affinity; D017931:DNA Primers; D000091342:Endoplasmic Reticulum Chaperone BiP; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000494:pharmacology*; D037161:Galectins / Q000494:pharmacology*; D006367:HeLa Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D012097:Repressor Proteins / Q000737:chemistry* / Q000494:pharmacology*; D014157:Transcription Factors / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014357:Trypsin
chemical_list: D017304:Annexin A5; D017931:DNA Primers; D000091342:Endoplasmic Reticulum Chaperone BiP; D005656:Fungal Proteins; D037161:Galectins; C000717247:HSPA5 protein, human; D037102:Lectins; C470833:MORF4L1 protein, human; D012097:Repressor Proteins; D014157:Transcription Factors; D014357:Trypsin