PMID- 35503584
OWN - NLM
STAT- MEDLINE
DCOM- 20220519
LR  - 20220531
IS  - 1573-8221 (Electronic)
IS  - 0007-4888 (Linking)
VI  - 172
IP  - 6
DP  - 2022 Apr
TI  - Identification and Verification of the Ability of Cdk5 to Phosphorylate 
      Deubiquitinating Enzyme BRCC3 In Vitro.
PG  - 701-708
LID - 10.1007/s10517-022-05460-z [doi]
AB  - It is known that the expression of the deubiquitinating enzyme 
      BRCA1-BRCA2-containing complex subunit 3 (BRCC3) and cyclin-dependent protein 
      kinase 5 (Cdk5) is increased in Parkinson's disease (both are involved in 
      neuroinflammatory response). However, the regulatory mechanism of Cdk5 on the 
      post-translational modification of BRCC3 remains unclear. Here we studied whether 
      Cdk5 phosphorylates BRCC3. Phosphorylation of BRCC3 by Cdk5 was predicted by GPS 
      5.0 software. His-BRCC3 plasmid was constructed by cloning the BRCC3 gene into 
      pGEX-6P-1 vector, and then His-BRCC3 fusion protein was induced with isopropyl 
      beta-d-1-thiogalactopyranoside and purified using His-Tag affinity chromatography 
      purification agarose. Phosphorylation of BRCC3 fusion protein by Cdk5 in vitro 
      was detected by mass spectrometry and Western blotting. The results showed that 
      multiple phosphorylation sites were predicted by GPS 5.0, and the His-BRCC3 
      fusion protein was successfully induced and purified. In vitro kinase assay, 
      Western blotting, and mass spectrometry showed that Cdk5 can phosphorylate BRCC3. 
      It has been demonstrated that protein kinase Cdk5 can phosphorylate the 
      deubiquitinating enzyme BRCC3 in vitro, which provides new data for further study 
      on the mechanism of neurodegeneration.
CI  - (c) 2022. Springer Science+Business Media, LLC, part of Springer Nature.
FAU - Zhang, C H
AU  - Zhang CH
AD  - Department of Human Anatomy, Laboratory Animal Center, Guilin Medical College, 
      Guilin, Guangxi, China.
FAU - Qin, K
AU  - Qin K
AD  - Department of Human Anatomy, Laboratory Animal Center, Guilin Medical College, 
      Guilin, Guangxi, China.
FAU - Tian, S L
AU  - Tian SL
AD  - Department of Human Anatomy, Laboratory Animal Center, Guilin Medical College, 
      Guilin, Guangxi, China.
FAU - Han, Y
AU  - Han Y
AD  - Department of Human Anatomy, Laboratory Animal Center, Guilin Medical College, 
      Guilin, Guangxi, China.
FAU - Xu, S Y
AU  - Xu SY
AD  - Laboratory Animal Center, Guilin Medical College, Guilin, Guangxi, China. 
      shaoyexu@glmc.edu.cn.
FAU - Shao, X Y
AU  - Shao XY
AD  - Department of Human Anatomy, Laboratory Animal Center, Guilin Medical College, 
      Guilin, Guangxi, China. xyshao@glmc.edu.cn.
LA  - eng
PT  - Journal Article
DEP - 20220503
PL  - United States
TA  - Bull Exp Biol Med
JT  - Bulletin of experimental biology and medicine
JID - 0372557
RN  - EC 2.7.11.1 (Cyclin-Dependent Kinase 5)
RN  - EC 2.7.11.22 (CDK5 protein, human)
RN  - EC 3.4.19.12 (BRCC3 protein, human)
RN  - EC 3.4.19.12 (Deubiquitinating Enzymes)
SB  - IM
MH  - Blotting, Western
MH  - *Cyclin-Dependent Kinase 5/metabolism
MH  - *Deubiquitinating Enzymes/genetics/metabolism
MH  - Humans
MH  - Parkinson Disease/metabolism
MH  - Phosphorylation
OTO - NOTNLM
OT  - BRCA1-BRCA2-containing complex subunit 3 (BRCC3)
OT  - cyclin-dependent kinase 5 (Cdk5)
OT  - kinase reaction
OT  - phosphorylation
OT  - phosphorylation mass spectrometry
EDAT- 2022/05/04 06:00
MHDA- 2022/05/20 06:00
CRDT- 2022/05/03 12:54
PHST- 2021/06/07 00:00 [received]
PHST- 2022/05/04 06:00 [pubmed]
PHST- 2022/05/20 06:00 [medline]
PHST- 2022/05/03 12:54 [entrez]
AID - 10.1007/s10517-022-05460-z [pii]
AID - 10.1007/s10517-022-05460-z [doi]
PST - ppublish
SO  - Bull Exp Biol Med. 2022 Apr;172(6):701-708. doi: 10.1007/s10517-022-05460-z. Epub 
      2022 May 3.