|Detection of Human and Mouse LRRC4 by Western Blot. Western blot shows lysates of human brain (hypothalamus) tissue and mouse brain tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human LRRC4 Monoclonal Antibody (Catalog # MAB4995) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for LRRC4 at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
LRRC4 (Leucine rich repeat/LRR-containing protein 4), also called NGL-2 (netrin-G ligand-2) or NAG14 (nasopharyngeal carcinoma-associated gene 14) is a 55 kDa (predicted, unglycosylated) type I transmembrane protein that is a member of the NGL family of synaptic LRR adhesion molecules (1, 2). Human LRRC4 cDNA encodes 653 amino acids (aa) that include a 38 aa signal sequence, a 489 aa extracellular domain (ECD), a 21 aa transmembrane domain, and a 105 aa cytoplasmic domain. The ECD contains nine LRRs (aa 74‑288), a C2 type Ig like domain (aa 354‑440), and a Thr-rich segment (aa 455‑526). Within the ECD, human LRRC4 shares 98% aa identity with mouse and rat, 99% aa identity with canine and bovine, and 99.6% aa identity with equine LRRC4. It also shares 54‑55% aa identity with family members LRRC4C/NGL-1 and LRRC4B/NGL-3, but each recognizes different ligands (1). LRRC4 is predominantly expressed in the brain on neurons and astrocytes as a ligand for netrin-G2 on the dendritic surface of synaptic neurons (2-4). It is proposed to regulate the formation of excitatory synapses via recruitment of PSD-95 to the cytoplasmic domain after aggregation of LRRC4 at the surface (3, 5). It suppresses proliferation by downregulating cell signaling pathways, resulting in altered expression of cell cycle regulating proteins and delay at the late G1 phase (1, 2, 6-8). It is thus considered a tumor suppressor protein and is often downregulated in brain tumors, particularly gliomas (1, 2, 6). Forced expression of LRRC4 in tumor cells slows proliferation and promotes differentiation (1, 4, 9). Addition of soluable LRRC4 to cultured neurons reduces excitatory synapse formation (3).
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