P4HB Human, Active

Prolyl 4-Hydroxylase Beta Human Recombinant, Active
Cat. No.
BT29289
Source
Escherichia Coli.
Synonyms
P4Hbeta, PDI, PDIA1, PHD, PO4DB, PO4HB, ERBA2L.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 90.0% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

P4HB Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 512 amino acids (18-508 a.a.) and having a molecular mass of 57.5kDa. The P4HB is fused to a 21 amino acid His Tag and purified by conventional chromatography.

Product Specs

Introduction
The enzyme P4HB, a member of the protein disulfide isomerase family, is both multifunctional and highly abundant. P4HB plays a role in hydroxylating prolyl residues within preprocollagen when found as a tetramer comprising two alpha and two beta subunits. This disulfide isomerase possesses two thioredoxin domains that facilitate the formation, breakage, and rearrangement of disulfide bonds.
Description
Recombinant human P4HB, produced in E. coli, is a single, non-glycosylated polypeptide chain consisting of 512 amino acids (specifically, amino acids 18-508). It has a molecular weight of 57.5 kDa. The P4HB protein is fused to a 21 amino acid His Tag for purification purposes, which is achieved using conventional chromatography techniques.
Physical Appearance
A clear, sterile-filtered solution.
Formulation
The P4HB protein is supplied at a concentration of 1 mg/ml in a solution containing 20 mM Tris-HCl at a pH of 8 and 10% glycerol.
Stability
For optimal storage, keep the vial at 4°C if it will be fully used within 2-4 weeks. For prolonged storage, it is recommended to freeze the product at -20°C. The addition of a carrier protein (either 0.1% HSA or BSA) is advised for long-term storage. To maintain product integrity, avoid repeated freeze-thaw cycles.
Purity
The purity of this product exceeds 90.0% as determined by SDS-PAGE analysis.
Biological Activity
The specific activity of this product is greater than 100 A650/cm/min/mg. Enzymatic activity was validated by assessing the aggregation of insulin in the presence of DTT.
Synonyms
P4Hbeta, PDI, PDIA1, PHD, PO4DB, PO4HB, ERBA2L.
Source
Escherichia Coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MDAPEEEDHV LVLRKSNFAE ALAAHKYLLV EFYAPWCGHC KALAPEYAKA AGKLKAEGSE IRLAKVDATE ESDLAQQYGV RGYPTIKFFR NGDTASPKEY TAGREADDIV NWLKKRTGPA ATTLPDGAAA ESLVESSEVA VIGFFKDVES DSAKQFLQAA EAIDDIPFGI TSNSDVFSKY QLDKDGVVLF KKFDEGRNNF EGEVTKENLL DFIKHNQLPL VIEFTEQTAP KIFGGEIKTH ILLFLPKSVS DYDGKLSNFK TAAESFKGKI LFIFIDSDHT DNQRILEFFG LKKEECPAVR LITLEEEMTK YKPESEELTA ERITEFCHRF LEGKIKPHLM SQELPEDWDK QPVKVLVGKN FEDVAFDEKK NVFVEFYAPW CGHCKQLAPI WDKLGETYKD HENIVIAKMD STANEVEAVK VHSFPTLKFF PASADRTVID YNGERTLDGF KKFLESGGQD GAGDDDDLED LEEAEEPDME EDDDQKAVKD EL.

Product Science Overview

Structure and Function

P4HB functions as a tetramer consisting of two alpha and two beta subunits . The beta subunit, encoded by the P4HB gene, is essential for the enzyme’s activity. The primary role of P4HB is to hydroxylate proline residues in collagen, a critical post-translational modification that stabilizes the collagen triple helix structure . This modification is vital for the proper formation and function of collagen, which is the most abundant protein in the extracellular matrix .

Recombinant Expression

Recombinant human P4HB is produced using various expression systems, including bacterial and yeast systems . These systems are engineered to express the P4HB gene, allowing for the production of the enzyme in large quantities. The recombinant form of P4HB retains its enzymatic activity and is used in various research and industrial applications .

Applications
  1. Collagen Production: P4HB is used in the production of recombinant collagen, which has applications in tissue engineering, regenerative medicine, and cosmetic products .
  2. Biochemical Research: The enzyme is used to study the biochemical and biophysical properties of collagen and other proteins that undergo proline hydroxylation .
  3. Clinical Diagnostics: P4HB is utilized in diagnostic assays to detect and quantify collagen and other hydroxylated proteins in biological samples .
Recent Advances

Recent studies have focused on optimizing the production and activity of recombinant P4HB. For example, a sandwich ELISA method has been developed for the specific and quantitative detection of active P4HB in crude samples . This method is highly sensitive and can be used to monitor P4HB levels during recombinant production processes .

Additionally, new liquid chromatography-mass spectrometry (LC-MS) techniques have been developed to assay proline hydroxylation in recombinant collagen variants . These techniques provide accurate and precise measurements of hydroxylation levels, which are critical for ensuring the quality and functionality of recombinant collagen .

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