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Hydroxyproline

Hydroxyproline

specific proline residues on the amino side of a glycine residue in collagen become hydroxylated at C4, before the polypeptides become helical, by the activity of prolyl hydroxylase. This enzyme has a ferrous ion at the active site and a reducing agent such as ascorbate is necessary to maintain the iron in the ferrous state. The presence of hydroxyproline is essential to produce stable triple helical tropocollagen, hence the problems caused by ascorbate deficiency in scurvy. This unusual amino acid is also present in considerable amounts in the major glycoprotein of primary plant cell walls (see HRGP).


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hydroxyproline

Insoluble cross-linked collagen can be measured by determination of the 4-hydroxyproline upon hydrolysis. There are now a few commercially available kits (Biovision and Quickzyme) to quantify the hydroxyproline or measure total collagen including acid hydrolysis ...

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by thematrix
Fri Dec 23, 2011 2:30 pm
 
Forum: Molecular Biology
Topic: hydroxyproline
Replies: 2
Views: 2029

Re: hydroxyproline assay to determine collagen content

Hi, This is a kit that works well :Catalog number K555-100 from Biovision. It's a colorimetric assay similar to yours. The standards work nicely, although the range could be extended if needed. Good luck ! E.

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by enndiaye
Fri Nov 04, 2011 4:24 pm
 
Forum: Molecular Biology
Topic: hydroxyproline assay to determine collagen content
Replies: 5
Views: 8259

hydroxyproline

... SPARC-null mice were studied. LV collagen content was measured histologically by collagen volume fraction, collagen composition was measured by hydroxyproline assay as soluble collagen (1 M NaCl extractable) versus insoluble collagen (mature cross-linked), and collagen morphological structure ...

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by HerrBiologie
Sun Oct 30, 2011 9:12 am
 
Forum: Molecular Biology
Topic: hydroxyproline
Replies: 2
Views: 2029

Re: hydroxyproline assay to determine collagen content

Hello Stip, I have an adjusted protocol for intervertebral discs. This assay estimates the amount of hydroxyproline, which has been shown to account for approximately 14% the amino acids in collagen. Materials: Hydroxyproline Standard - 1-5 µg/2 0.001N HCl ml. Buffer-0.5 ...

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by ucsfmed2012
Thu Oct 27, 2011 9:30 am
 
Forum: Molecular Biology
Topic: hydroxyproline assay to determine collagen content
Replies: 5
Views: 8259

hydroxyproline

Can the 4-hyydroxyproline measure insoluble crosslink collagen content in heart tissue?

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by blazing
Tue Oct 18, 2011 6:53 am
 
Forum: Molecular Biology
Topic: hydroxyproline
Replies: 2
Views: 2029
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