Identification of Microprotein-Protein Interactions via APEX Tagging
被引:46
作者:
Chu, Qian
论文数: 0引用数: 0
h-index: 0
机构:
Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USASalk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Chu, Qian
[1
]
Rathore, Annie
论文数: 0引用数: 0
h-index: 0
机构:
Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Univ Calif San Diego, Div Biol Sci, 9500 Gilman Dr, La Jolla, CA 92093 USASalk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Rathore, Annie
[1
,2
]
Diedrich, Jolene K.
论文数: 0引用数: 0
h-index: 0
机构:
Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Scripps Res Inst, Dept Physiol Chem, 10550 North Torrey Pines Rd, La Jolla, CA 92037 USASalk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Diedrich, Jolene K.
[1
,3
]
Donaldson, Cynthia J.
论文数: 0引用数: 0
h-index: 0
机构:
Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USASalk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Donaldson, Cynthia J.
[1
]
Yates, John R., III
论文数: 0引用数: 0
h-index: 0
机构:
Scripps Res Inst, Dept Physiol Chem, 10550 North Torrey Pines Rd, La Jolla, CA 92037 USASalk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Yates, John R., III
[3
]
Saghatelian, Alan
论文数: 0引用数: 0
h-index: 0
机构:
Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USASalk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
Saghatelian, Alan
[1
]
机构:
[1] Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, 10010 North Torrey Pines Rd, La Jolla, CA 92037 USA
[2] Univ Calif San Diego, Div Biol Sci, 9500 Gilman Dr, La Jolla, CA 92093 USA
[3] Scripps Res Inst, Dept Physiol Chem, 10550 North Torrey Pines Rd, La Jolla, CA 92037 USA
OPEN READING FRAMES;
LIVING CELLS;
MESSENGER-RNA;
PEPTIDES;
NUCLEOPHOSMIN;
DISCOVERY;
SIGNAL;
POLYPEPTIDES;
DROSOPHILA;
NETWORK;
D O I:
10.1021/acs.biochem.7b00265
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
Microproteins are peptides and small proteins encoded by small open reading frames (smORFs). Newer technologies have led to the recent discovery of hundreds to thousands of new microproteins. The biological functions of a few inicropioteins have been elucidated, and these micro-proteins have fundamental roles in biology ranging from limb development to muscle function, highlighting the Value of characterizing these molecules. The identification of microprotein protein interactions,(MPIs) has proven to be a successful approach to the functional characterization of these genes; however, traditional immunoprecipitation methods result in the, enrichment of nonspecific. interactions for, microproteins. Here, we test and apply an in situ proximity tagging method that relies on an engineered ascorbate peroxidase 2 (APEX) to elucidate MPls. The results,demonstrate that APEX tagging is superior. to traditional immunoprecipitation methods for microproteins. Furthermore,the application of APEX tagging to an uncharacterized microprotein called Cllorf98 revealed that this microprotein interacts with nucleolar proteins nucleophosmin and nucleolin, demonstrating the ability,of this approach to identify novel. hypothesis-generating MPIs.