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噬菌体λ编码的一种蛋白质丝氨酸/苏氨酸磷酸酶的表达及生化特性

Expression and biochemical properties of a protein serine/threonine phosphatase encoded by bacteriophage lambda.

作者信息

Barik S

机构信息

Department of Molecular Biology, Cleveland Clinic Foundation, OH 44195.

出版信息

Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10633-7. doi: 10.1073/pnas.90.22.10633.

DOI:10.1073/pnas.90.22.10633
PMID:8248155
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC47831/
Abstract

The predicted amino acid sequence encoded by the open reading frame 221 (orf221) of bacteriophage lambda exhibited a high degree of similarity to the catalytic subunits of a variety of protein serine/threonine phosphatases belonging to PP1, PP2A, and PP2B groups. Cloning and expression of the orf221 gene in Escherichia coli provided direct evidence that the gene codes for a protein serine/threonine phosphatase. The single-subunit recombinant enzyme was purified in soluble form and shown to possess a unique repertoire of biochemical properties--e.g., an absolute requirement for Mn2+, resistance to okadaic acid, inhibitors 1 and 2, and ability to dephosphorylate casein, adenovirus E1A proteins, and the alpha subunit of phosphorylase kinase. No phosphotyrosine phosphatase activity was observed. Mutational and biochemical analyses identified the conserved residues 73-77 and Cys138 to be important for activity. The name PP-lambda is proposed for this unusual prokaryotic enzyme.

摘要

噬菌体λ开放阅读框221（orf221）编码的预测氨基酸序列与属于PP1、PP2A和PP2B组的多种蛋白质丝氨酸/苏氨酸磷酸酶的催化亚基具有高度相似性。orf221基因在大肠杆菌中的克隆和表达提供了直接证据，证明该基因编码一种蛋白质丝氨酸/苏氨酸磷酸酶。单亚基重组酶以可溶形式纯化，并显示出具有独特的生化特性——例如，绝对需要Mn2+，对冈田酸、抑制剂1和2有抗性，以及能够使酪蛋白、腺病毒E1A蛋白和磷酸化酶激酶的α亚基去磷酸化。未观察到磷酸酪氨酸磷酸酶活性。突变和生化分析确定保守残基73 - 77和Cys138对活性很重要。为此种异常的原核酶提议命名为PP-λ。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/27f8b40c19a3/pnas01529-0234-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/0c415ddbf309/pnas01529-0232-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/38379c209181/pnas01529-0233-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/2e65c4d63b87/pnas01529-0233-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/eb677758f5ce/pnas01529-0234-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/27f8b40c19a3/pnas01529-0234-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/0c415ddbf309/pnas01529-0232-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/38379c209181/pnas01529-0233-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/2e65c4d63b87/pnas01529-0233-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/eb677758f5ce/pnas01529-0234-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9588/47831/27f8b40c19a3/pnas01529-0234-b.jpg

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