CD45, CD148, and Lyp/Pep: critical phosphatases regulating Src family kinase signaling networks in immune cells

Abstract

Reciprocal regulation of tyrosine phosphorylation by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs) is central to normal immune cell function. Disruption of the equilibrium between PTK and PTP activity can result in immunodeficiency, autoimmunity, or malignancy. Src family kinases (SFKs) play a central role in both immune cell function and disease due to their proximal position in numerous signal transduction cascades including those emanating from integrin, T and B-cell antigen receptors, Fc, growth factor, and cytokine receptors. Given that tight regulation of SFKs activity is critical for appropriate responses to stimulation of these various signaling pathways, it is perhaps not surprising that multiple PTPs are involved in their regulation. Here, we focus on the role of three phosphatases, CD45, CD148, and LYP/PEP, which are critical regulators of SFKs in hematopoietic cells. We review our current understanding of their structures, expression, functions in different hematopoietic cell subsets, regulation, and putative roles in disease. Finally, we discuss remaining questions that must be addressed if we are to have a clearer understanding of the coordinated regulation of tyrosine phosphorylation and signaling networks in hematopoietic cells and how they could potentially be manipulated therapeutically in disease.

Figure 1. SFKs are in a dynamic equilibrium between closed and active conformations

Interaction of the SH2 domain and the C-terminal negative regulatory phosphotyrosine results in a closed conformation with decreased access to the kinase's catalytic site. Dephosphorylation of this site results in adoption of a more open, or ‘primed’, conformation. This conformation can potentially also be facilitated when the relatively weak interaction between the SH2 domain and the C-terminal phosphotyrosine is overcome by competing interactions of other proteins (denoted in the schematic as X or Y) with the SH2 or SH3 domains of the SFK. Phosphorylation of the catalytic site tyrosine is required for full kinase activity. (Adapted from (2,7)).

Figure 2. CD45, CD148, and Lyp/Pep structures

Key functional domains are depicted. Both CD45 and CD148 are RPTPs while Lyp/Pep lacks a transmembrane domain and is cytoplasmic. CD45 exists as multiple isoforms due to alternative splicing of exons 4, 5, and 6 that encode three regions, designated A, B, and C, in the extracellular domain, which contain multiple sites for O-linked glycosylation and sialyation. The largest (RABC) and smallest (RO) isoforms are shown. While CD45 contains two PTP domains (of which only D1 is active), CD148 and Lyp/Pep contain only a single PTP domain.

Figure 3. Reciprocal regulation of SFK activation by CD45, CD148, and Lyp/Pep

SFKs are in a dynamic equilibrium between their inactive and active conformations in both the basal state and during immune cell activation (while B cell activation is depicted, similar mechanisms likely exist in other hematopoietic cell lineages). In the basal state, phosphorylation of the adapter PAG facilitates recruitment of Csk and Lyp/Pep. Csk phosphorylates the negative regulatory tyrosine while Lyp/Pep dephosphorylates the autocatalytic tyrosine. This results in the SFK adopting a closed conformation. By opposing Csk and dephosphorylating the negative regulatory tyrosine, CD45 and CD148 generate a pool of SFKs in an open or ‘primed’ conformation that can be rapidly activated should antigen be encountered. We hypothesize that low levels of SFK transphosphorylation of the autocatalytic site occur in the basal state, which is necessary for tonic signaling and cell survival. This is balanced by the action of PAG/Csk/Pep (and potentially CD45 targeting either directly or indirectly the autocatalytic tyrosine). During antigen encounter and receptor activation, PAG becomes dephosphorylated via mechanisms that are incompletely elucidated (but may involve CD45), Csk and Lyp/Pep dissociate from the cell surface leaving CD45 and CD148 actions on the negative regulatory tyrosine unopposed. Phosphorylation of the autocatalytic tyrosine is now favored. The fully active SFK can phosphorylate the ITAMs of the immunoreceptor, facilitating recruitment of Syk/Zap70 and amplification of downstream signaling cascades.