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  4. PAM Protein, Human (HEK293, His)

PAM Protein, Human (HEK293, His)

Cat. No.: HY-P73731
COA Handling Instructions

The PAM protein is a bifunctional enzyme that coordinates the α-amidation process, which is critical for the biosynthesis of neuropeptides and endocrine peptides. The peptidyl α-hydroxylating monooxygenase (PHM) domain hydroxylates the C-terminal glycine, and the peptidylglycine amide glycolate lyase (PAL) domain cleaves the NC-α bond, producing α-amidated peptides. PAM Protein, Human (HEK293, His) is the recombinant human-derived PAM protein, expressed by HEK293 , with C-His labeled tag.

For research use only. We do not sell to patients.

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  • Biological Activity

  • Technical Parameters

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Description

The PAM protein is a bifunctional enzyme that coordinates the α-amidation process, which is critical for the biosynthesis of neuropeptides and endocrine peptides. The peptidyl α-hydroxylating monooxygenase (PHM) domain hydroxylates the C-terminal glycine, and the peptidylglycine amide glycolate lyase (PAL) domain cleaves the NC-α bond, producing α-amidated peptides. PAM Protein, Human (HEK293, His) is the recombinant human-derived PAM protein, expressed by HEK293 , with C-His labeled tag.

Background

The PAM protein operates as a bifunctional enzyme, overseeing the post-translational modification of inactive peptidylglycine precursors to their bioactive alpha-amidated peptide forms—a crucial terminal modification in the biosynthesis of numerous neural and endocrine peptides. The alpha-amidation process involves two sequential reactions, each governed by distinct catalytic domains within the enzyme. In the first step, the peptidyl alpha-hydroxylating monooxygenase (PHM) domain catalyzes a copper-, ascorbate-, and O2-dependent stereospecific hydroxylation (with S stereochemistry) at the alpha-carbon (C-alpha) of the C-terminal glycine of the peptidylglycine substrate. The subsequent step, orchestrated by the peptidylglycine amidoglycolate lyase (PAL) domain, entails a zinc-dependent cleavage of the N-C-alpha bond, resulting in the production of the alpha-amidated peptide and glyoxylate. Additionally, PAM exhibits a similar capacity to catalyze the two-step conversion of an N-fatty acylglycine to a primary fatty acid amide and glyoxylate in a manner reminiscent of its peptidylglycine modification function.

Biological Activity

Measured by its ability to convert Hippurate to Benzamide and Glyoxylate. The specific activity is 731.87 pmol/min/μg, as measured under the described conditions.

Species

Human

Source

HEK293

Tag

C-His

Accession

P19021-2/NP_620176.1 (F21-S866)

Gene ID
Molecular Construction
N-term
PAM (F21-S866)
Accession # P19021-2/NP_620176.1
His
C-term
Synonyms
Peptidyl-glycine alpha-amidating monooxygenase; PAM; PHM; PAL
Molecular Weight

Approximately 94.4 kDa

Purity

Greater than 90% as determined by reducing SDS-PAGE.

Appearance

Lyophilized powder

Formulation

Lyophilized from a 0.2 μm filtered solution of PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween 80 are added as protectants before lyophilization.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.

Storage & Stability

Stored at -20°C for 2 years. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

Documentation

PAM Protein, Human (HEK293, His) Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

  • Reconstitution Calculator

  • Dilution Calculator

  • Specific Activity Calculator

The reconstitution calculator equation

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration
= ÷

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

The specific activity calculator equation

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)
Unit/mg = 106 ÷ ng/mL

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PAM Protein, Human (HEK293, His)
Cat. No.:
HY-P73731
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