What is a hydroxyproline assay?
Hydroxyproline assays generally involve reactions with oxidized hydroxyproline to produce a change in color, spectroscopically measured at an absorbance peak at 560 nm. Visit the hydroxyproline assay kit suppliers for more details, including assay method and included reagents.
How is hydroxyproline measured?
The measurement of hydroxyproline in plasma, urine and body tissue is possible by colorimetric methods, high-performance liquid chromatography (HPLC) and flow injection analyses (4–6). However, these methods require large sample volumes, due to their low sensitivity.
How do you calculate hydroxyproline collagen?
Convert hydroxyproline to collagen by multiplying the supernatants by 7.52 (Cross et al., 1973) and the residuals by 7.25 to μg collagen/g of sample.
Why is hydroxyproline important in collagen?
Collagen. Hydroxyproline is a major component of the protein collagen, comprising roughly 13.5% of mammalian collagen. Hydroxyproline and proline play key roles for collagen stability. They permit the sharp twisting of the collagen helix.
Is hydroxyproline an amino acid?
Proline and its metabolite (hydroxyproline) are unique amino acids (AA) both chemically and biochemically (Hu et al. 2008; Kaul et al. 2008). They constitute one-third of AA in the collagen proteins which comprise approximately 30% of body proteins.
What is urinary hydroxyproline?
Hydroxyproline, a metabolite of proline, which constitutes ~14% of the total amino acid content of collagen protein, has historically been measured in urine to measure collagen turnover and/or perturbations to collagen metabolism (Prockop et al., 1961).
What is hydroxyproline used for?
Hydroxyproline is mostly used as a diagnostic marker of bone turnover and liver fibrosis. Therapeutically, hydroxyproline is being studied as an an experimental medicine but is approved in France as a combination topical gel product called Cicactive for small, superficial wounds.
What is hydroxyproline and hydroxylysine?
Hydroxylysine is another amino acid unique to collagen and proteins containing collagen-like sequences. Like hydroxyproline, hydroxylysine is not re-utilized for collagen biosynthesis, and although it is much less abundant than hydroxyproline, it is a potential marker of collagen degradation.
What is the function of hydroxyproline?
Hydroxyproline is a major component of the protein collagen. Hydroxyproline and proline play key roles for collagen stability. They permit the sharp twisting of the collagen helix. It helps provide stability to the triple-helical structure of collagen by forming hydrogen bonds.
What is hydroxyproline composed of?
Hydroxyproline is formed from proline through hydroxylation of proline in proteins, primarily collagen (Wu et al., 2017c). Hydroxyproline is crucial for collagen synthesis.
What is hydroxyproline content?
Hydroxyproline is a major component of fibrillar collagen of all types, comprising ~14% of the total amino and imino acid content. It is produced by the post-translational modification of proline by the enzyme 4-prolyl hydroxylase. In plasma, hydroxyproline exists in protein-bound, peptide-bound and free forms.
What is hydroxylproline in collagen?
Hydroxyproline is a major component of collagen, where it serves to stabilize the helical structure. Because hydroxyproline is largely restricted to collagen, the measurement of hydroxylproline levels can be used as an indicator of collagen content.
How does the interstitial collagen assay complement the hydroxyproline assay?
This technique complements the hydroxyproline assay by providing a means of identifying which interstitial collagens are altered in renal disease. Publication types Research Support, Non-U.S. Gov’t
How do you test for collagen without hydroxyproline?
Therefore, without hydroxyproline, no functional collagen fibers appear in the extracellular space 3). Hydroxyproline can be tested in the serum and in the urine. The serum level of hydroxyproline excretion correlates well with its urinary level 4).
How is hydroxyproline concentration determined in the hydroxyproline assay kit?
In the Hydroxyproline Assay Kit, hydroxyproline concentration is determined by the reaction of oxidized hydroxyproline with 4- (Dimethylamino)benzaldehyde (DMAB), which results in a colorimetric (560 nm) product, proportional to the hydroxyproline present.