Starter Guides you to SHP Substrate 1 Recombinant Protein

Published by ANT BIO PTE. LTD.

1. Concept

SHP Substrate 1 (also known as SHPS1/SIRPα) is a key immune regulatory transmembrane protein that functions as a primary substrate for SHP‑1 and SHP‑2 phosphatases. It contains critical ITIM (Y241) and ITSM (Y263) motifs that mediate downstream inhibitory signaling, making it essential for studying immune tolerance, macrophage phagocytosis, T‑cell regulation, and cancer immune evasion. SHP Substrate 1 recombinant protein is a bioactive, genetically engineered tool widely used for molecular interaction, structural biology, drug screening, and diagnostic research.

2. Research Frontiers

Recent advances in SHP Substrate 1 recombinant protein technology:

• Rational design of truncated intracellular domains (183–268) that preserve full SHP‑1/SHP‑2 binding capacity.
• Use of flexible linkers to improve solubility, stability, and crystallization success.
• Multi‑system expression (bacterial, insect, mammalian) tailored for post‑translational modifications.
• High‑resolution structural analysis of SHP Substrate 1–SHP‑2 complexes via X‑ray crystallography and cryo‑EM.
• Application in high‑throughput drug screening, CAR‑T engineering, and clinical diagnostic development.

3. Research Significance

SHP Substrate 1 recombinant protein is a cornerstone for immune and signaling research:

• Reveals the molecular mechanism of SHP‑1/SHP‑2 activation and immune checkpoint regulation.
• Enables structure‑based drug design for cancer, autoimmunity, and inflammation.
• Supports quantitative detection and biomarker development in clinical samples.
• Facilitates cell therapy optimization and gene editing validation.

4. Molecular Design, Expression, and Functional Applications

4.1 Molecular Design & Expression Systems

High‑performance SHP Substrate 1 recombinant protein is often designed as a truncated intracellular fragment (residues 183–268) containing functional ITIM and ITSM motifs. Common tags include GST, His6, FLAG, and HA to support purification and detection. Flexible 15–20 aa linkers enhance solubility and crystallization.

Three expression systems are widely used:

• E. coli: High yield (50–100 mg/L), low cost; ideal for non‑glycosylated protein.
• Insect cells (Sf9): Supports palmitoylation and glycosylation.
• Mammalian cells (HEK293F): Produces native‑like protein with highest binding affinity (Kd ≈ 0.3 μM).

[Image Location: Schematic diagram of SHP Substrate 1 – SHP‑2 interaction and domain organization]

4.2 Purification & Quality Control

Purification uses affinity and size‑exclusion chromatography to achieve >95% purity with endotoxin levels <0.1 EU/μg. Key quality assessments include:

• SDS‑PAGE and HPLC for purity
• Mass spectrometry for molecular weight and modifications
• SPR for binding affinity to SHP‑2 (Kd = 0.5–2 μM)
• Dynamic light scattering (DLS) for monodispersity
• Stability testing at −80°C for long‑term storage

4.3 Protein Modification & Functional Validation

Phosphorylation at Y241 and Y263 activates SHP‑2 up to 100‑fold. Dual phosphorylation shows synergistic signaling enhancement. Palmitoylation and glycosylation support membrane localization and physiological function. Validated assays include:

• In vitro phosphatase activity measurement
• SPR binding kinetics
• TCR signaling inhibition in cell models
• Batch‑to‑batch consistency control

4.4 Molecular Interactions & Structural Biology

SHP Substrate 1 acts as a signaling hub interacting with SHP‑1, SHP‑2, Grb2, 14‑3‑3, Cbl, and kinases. Structural studies (PDB: 6N6F; EMDB: 3067) reveal how dual phosphorylated motifs trigger SHP‑2 conformational activation, opening the catalytic cleft by ~15°. These insights enable rational drug design targeting the SHP Substrate 1–SHP‑2 axis.

4.5 Research & Translational Applications

• Drug Screening: HTS for small‑molecule modulators of SHP‑2 activity.
• Diagnostics: ELISA and flow cytometry standards for soluble SHP Substrate 1 detection.
• Cell Therapy: Improves CAR‑T persistence and Treg differentiation.
• Basic Research: In vitro reconstitution of inhibitory signalosomes and single‑molecule dynamics.

5. Our Mission at ANT BIO PTE. LTD.

ANT BIO PTE. LTD. empowers breakthroughs in immunology, oncology, and drug discovery by providing highly active, consistent, and well‑validated SHP Substrate 1 recombinant proteins and supporting reagents. Our products enable reliable results in molecular interaction, signaling, screening, and translational research.

6. Featured SHP Substrate 1–Related Products

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