Expression of LY6D is induced at the surface of MCF10A cells by X-ray irradiation.
In order to identify membrane proteins whose expression is induced by X-ray irradiation, we developed an antibody (Ab)-directed strategy using a phage Ab library. X-Ray-irradiated cells were screened with a phage Ab library in the presence of a large excess of polyclonal Abs prepared against membrane proteins that are commonly present at the surface of both X-ray-irradiated and nonirradiated cells. After isolation of Ab that bound only to X-ray-irradiated cells, the antigen was identified using MS. Using this approach, we found that expression of LY6D is induced in MCF10A cells by X-ray irradiation. The induction of LY6D expression is triggered through a pathway regulated by ATM, CHK2 and p53. This method is a new Ab-directed proteomic strategy for analysis of membrane proteins, and is applicable to various biological phenomena in situations in which both target molecule-expressing cells and nonexpressing cells are available.
Department of Biochemistry, Nagoya University School of Medicine, Japan.
Maiko
M
Kurosawa
Anand D
AD
Jeyasekharan
Eva-Maria
EM
Surmann
Noboru
N
Hashimoto
Vignesh
V
Venkatraman
Gene
G
Kurosawa
Koichi
K
Furukawa
Ashok R
AR
Venkitaraman
Yoshikazu
Y
Kurosawa
D064007:Ataxia Telangiectasia Mutated Proteins; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D064447:Checkpoint Kinase 2; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D058851:GPI-Linked Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D006801:Humans; D009153:Mutagens / Q000633:toxicity; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D034741:RNA, Small Interfering; D060888:Real-Time Polymerase Chain Reaction; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D014965:X-Rays
D015815:Cell Adhesion Molecules; D018797:Cell Cycle Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D058851:GPI-Linked Proteins; C094237:LY6D protein, human; D009153:Mutagens; D034741:RNA, Small Interfering; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D064447:Checkpoint Kinase 2; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C578634:CHEK2 protein, human; D017346:Protein Serine-Threonine Kinases; D016283:Proto-Oncogene Proteins p21(ras)