Identification of  L-lactate Dehydrogenase as a Protein Tyrosine Phosphatase 1B substrate using K-BIPS.

Identification of L-lactate Dehydrogenase as a Protein Tyrosine Phosphatase 1B substrate using K-BIPS. Chembiochem. 2020 Oct 01;: Authors: Acharige NPN, Pflum MKH Abstract Kinases and phosphatases are major players in a variety of cellular events, including cell signaling. Aberrant activity or mutations in kinases and phosphatases can lead to diseases, such as cancer, diabetes, and Alzheimer's. Compared to kinases, phosphatases are understudied, which is partly a result of the limited methods to identify substrates. As a solution, we developed a proteomics-based method called Kinase-catalyzed Biotinylation to Identify Phosphatase Substrates (K-BIPS) that previously identified substrates of Ser/Thr phosphatases using small molecule inhibitors. Here, for the first time, K-BIPS was applied to identify substrates of a tyrosine phosphatase, Protein Tyrosine Phosphatase 1B (PTP1B), using siRNA knockdown conditions. Eight possible substrates of PTP1B were discovered in HEK293 cells, including the known substrate Pyruvate Kinase. In addition, L-lactate Dehydrogenase (LDHA) was validated as a novel PTP1B substrate. With the ability to use knockdown conditions with Ser/Thr or Tyr phosphatases, K-BIPS represents a general discovery tool to explore phosphatases biology by identifying unanticipated substrates. PMID: 33002308 [PubMed - as supplied by publisher]
Source: Chembiochem - Category: Biochemistry Authors: Tags: Chembiochem Source Type: research