Class III PI 3-kinase is a subgroup of the enzyme family, phosphoinositide 3-kinase that share a common protein domain structure, substrate specificity and method of activation. There is only one known class III PI 3-kinase, Vps34, which is also the only PI 3-kinase expressed in all eukaryotic cells. In humans it is encoded by the PIK3C3 gene. In human cells Vps34 associates with a regulatory subunit, PIK3R4(p150, Vps15). Vps34 was first identified in a Saccharomyces cerevisiae (budding yeast) screen for proteins involved vesicle-mediated vacuolar protein sorting (hence Vps). A number of proteins containing a phosphoinositide binding domain specific for PtdIns(3)P that function in cellular protein trafficking have been identified.
背景文献
1. Jia M et al. ULK1-mediated metabolic reprogramming regulates Vps34 lipid kinase activity by its lactylation. Sci Adv. 2023 Jun
2. Liu Y et al. Targeting VPS34 in autophagy: An update on pharmacological small-molecule compounds. Eur J Med Chem. 2023 Aug
亚细胞定位
Midbody, Late endosome, Cytoplasmic vesicle, autophagosome.
Western blot analysis of VPS34 on different lysates with Rabbit anti-VPS34 antibody (HA723229) at 1/5,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: PC-3 cell lysate (20 µg/Lane) Lane 3: C2C12 cell lysate (20 µg/Lane) Lane 4: C6 cell lysate (20 µg/Lane) Lane 5: Mouse heart tissue lysate (40 µg/Lane) Lane 6: Mouse brain tissue lysate (40 µg/Lane) Lane 7: Rat heart tissue lysate (40 µg/Lane)
Predicted band size: 102 kDa Observed band size: 98 kDa
Exposure time: 15 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723229) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of VPS34 on different lysates with Rabbit anti-VPS34 antibody (HA723229) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-VPS34 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 102 kDa Observed band size: 98 kDa
Exposure time: 4 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723229) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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