Analysis of Protein Palmitoylation by Metabolic Radiolabeling Methods-猫眼课题宝

通过代谢放射性标记方法分析蛋白质棕榈酰化

DOI：

10.1007/978-1-59745-198-7_166

发表时间：

2009

期刊：

The Biochemical journal

影响因子：

通讯作者：

中科院分区：

文献类型：

作者：

Katherine H. Pedone;L. Bernstein;M. Linder;J. Hepler

Many eukaryotic signaling proteins are modified by the covalent attachment of long-chain lipids. These highly hydrophobic molecules bind target proteins and facilitate interaction with cellular membranes, lipid molecules and other proteins. Generally, there are three classes of lipids that modify target proteins: myristate, isoprenoids and palmitate (for reviews, see refs. 1,2). Myristate, a 14-carbon saturated fatty acid, binds proteins at N-terminal glycine residues via an amide linkage. This usually occurs cotranslationally after removal of the initiating methionine, although it also can occur after proteolytic cleavage and exposure of an internal glycine. Long-chain isoprenoid lipids, including farnesyl and geranylgeranyl groups, modify proteins posttranslationally and attach via a thioether linkage to C-terminal cysteine residues. Palmitate is a 16-carbon saturated fatty acid that modifies proteins posttranslationally through thioester incorporation at cysteines. Palmitate also can modify proteins at additional sites and by alternative mechanisms, as in proteins palmitoylated at serine and threonine residues via oxyester linkages and by amide-linked N-terminal cysteines and glycines. In addition, other fatty acid species form thioester linkages with proteins. For these reasons, the general term for protein lipid modification by thioester attachment is thioacylation, or S-acylation, and the term palmitoylation refers specifically to protein modification by palmitate. Proteins that undergo palmitoylation by thioester attachment at cysteine residues, or S-palmitoylation, represent the majority of protein targets of palmitoylation. This chapter will focus on methods for identifying protein substrates for thioester incorporation of palmitate.