ABSTRACT
The tubulin tyrosinylation/detyrosinylation cycle is a well-established posttranslational modification, which is carried out by two enzymes: tubulin tyrosine ligase (TTL) and tubulin tyrosine carboxypeptidase (TTCP). In this paper, I present evidence suggesting that the cycle itself is under the hierarchical control of reversible phosphorylation and that proteinkinase C (PKC)-mediated phosphorylation of TTL inhibits its activity, thereby preventing tubulin tyrosinylation. Phosphorylation of TTL is postulated to occur in its presumed Mg(++)-ATP binding fold, leading to inhibition of Mg(++)/ATP binding and TTL mediated catalysis. The implications of such control are also discussed. Copyright 2001 Harcourt Publishers Ltd.
ABSTRACT
Tumour Necrosis Factor alpha (TNF alpha), is an inflammatory cytokine produced by macrophages/monocytes during acute inflammation and is responsible for a diverse range of signalling events within cells, leading to necrosis or apoptosis. The protein is also important for resistance to infection and cancers. TNF alpha exerts many of its effects by binding, as a trimer, to either a 55 kDa cell membrane receptor termed TNFR-1 or a 75 kDa cell membrane receptor termed TNFR-2. Both these receptors belong to the so-called TNF receptor superfamily. The superfamily includes FAS, CD40, CD27, and RANK. The defining trait of these receptors is an extra cellular domain comprised of two to six repeats of cysteine rich motifs. Additionally, a number of structurally related "decoy receptors" exist that act to sequester TNF molecules, thereby rescuing cells from apoptosis. The crystal structures of TNF alpha, TNF beta, the extracellular domain of TNFR-1 (denoted sTNFR-1), and the TNF beta sTNFR-1 complex have been defined by crystallography. This article will review the structure/function relationships of the TNF alpha and the TNF receptor superfamily. It will also discuss insights as to how structural features play a role in the pleiotropic effects of TNF alpha.
Subject(s)
Receptors, Tumor Necrosis Factor/chemistry , Tumor Necrosis Factor-alpha/chemistry , Amino Acid Sequence , Animals , Humans , Molecular Sequence Data , Receptors, Tumor Necrosis Factor/metabolism , Structure-Activity Relationship , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/physiologyABSTRACT
The tubulin tyrosination/detyrosination cycle is a well-established posttranslational modification, which is carried out by two enzymes: Tubulin Tyrosine Ligase (TTL) and Tubulin Tyrosine Carboxypeptidase (TTCP). In this paper, I present evidence suggesting that the cycle itself is under the hierarchical control of reversible phosphorylation and that PKC mediated phosphorylation of TTL inhibits its activity, thereby preventing tubulin tyrosination. Phosphorylation of TTL is predicted to occur in a postulated Mg(++)/-ATP binding fold, leading to inhibition of Mg(++)/ATP binding and TTL mediated catalysis. The implications of such control are also discussed.
Subject(s)
Microtubules/enzymology , Peptide Synthases/metabolism , Tubulin/metabolism , Tyrosine/metabolism , Amino Acid Sequence , Animals , Humans , Molecular Sequence Data , Peptide Synthases/genetics , PhosphorylationABSTRACT
P-glycoprotein (Pgp) is a transmembrane transporter causing efflux of a number of chemically unrelated drugs and is responsible for resistance to a variety of anticancer drugs during chemotherapy. Pgp overexpression in cells is also associated with volume-activated chloride channel activity; Pgp is thought to regulate such activity. Reversible phosphorylation is a possible mechanism for regulating the transport and chloride channel regulation functions of Pgp. Protein kinase C (PKC) is a good candidate for inducing such phosphorylation. Hierarchical multiple phosphorylation (e.g. of different serines and with different PKC isoforms) may shuttle the protein between its different states of activity (transport or channel regulation). Cell volume changes may trigger phosphorylation of Pgp at sites causing inhibition of transport. The possible regulation of chloride channels by Pgp and the potential involvement of reversible phosphorylation in such regulation is reviewed.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Chloride Channels/metabolism , Eukaryotic Cells/metabolism , Ion Channel Gating/physiology , Animals , Cell Size/physiology , Eukaryotic Cells/cytology , Humans , PhosphorylationABSTRACT
TNF-alpha induces tumor-selective cytotoxicity in certain cancers, but many tumors are resistant to the effects of this inflammatory cytokine. This brief hypothesis outlines my views that nitric oxide-mediated alpha-tubulin posttranslational nitrotyrosination may be a major mechanism through which TNF-alpha exerts its cytotoxic effects on cancer cells. Additionally, I propose that tumor cells that are resistant to the effects of TNF-alpha may be so because of suppressed levels of "tubulin tyrosine ligase," which is responsible for the posttranslational tyrosination of alpha-tubulin.
