Observations on the gel chromatography of collagen-like peptides and use of a simple calibration method for molecular weight determination
✍ Scribed by N. Venkateswara Rao; Elijah Adams
- Publisher
- Elsevier Science
- Year
- 1975
- Tongue
- English
- Weight
- 507 KB
- Volume
- 68
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Elution characteristics of collagen-derived polypeptides and of globular proteins were compared under identical experimental conditions with agarose gels. This comparison permitted calculation of the hydrodynamic radii of collagen polypeptide chains of different molecular weight, and these radii were shown to be in reasonable agreement with estimates made from intrinsic viscosity data. Two distinct linear relationships were observed for collagen polypeptide chains, relating logarithm of molecular weight to elution parameters. Peptide chains of MW 3300 and lower fell on a line of a steeper slope than did larger polypeptide chains.
A simple procedure for molecular weight estimation of an unknown polypeptide chain of the collagen class is described, using only three commercially available standards for calibration: reduced, carboxymethylated Ascaris cuticle collagen, and the synthetic peptides (L-Pro-L-Pro-Gly)~0 and (L-Pro-L-Pro-Gly) 5.
Gel filtration in denaturing solvents (1,2) and gel electrophoresis (3) in sodium dodecyl sulfate I have been used with peptides derived from collagen. For relatively large polypeptides, these methods have yielded satisfactory straight-line relationships between logarithm of molecular weight and elution or mobility parameters. Our own need for a simple method both of estimating the molecular weight of collagen-like synthetic polypeptides and of fractionating them preparatively by size, led us to investigate gel filtration in somewhat greater detail than in past studies, particularly for the lower molecular weight range of 1000-10,000, for which sedimentation methods are rather unreliable.
Our findings extend earlier reports on collagen-like peptides (1,2), providing a systematic comparison of the elution behavior of globular proteins and collagen-like polypeptides, a comparison which has been