Thermal stability of calf skin collagen type I in salt solutions

The effects of guanidinium salts in decreasing the renaturation rate and lowering the thermal stability of acid-soluble calf-skin collagen have been compared with those of formamide and urea. With the exception of guanidinium sulphate at higher concentrations, no qualitative differences were apparent in the effects of these perturbants, which thus differed only in molar activity. Activity variation in the guanidinium salts reflected a net effect resulting from additivity of cation and anion contributions. As observed in other protein systems, lyotropic activity increased in the series formamide<urea<guanidinium ion, and in the guanidinium salts in the anion order fluoride<sulphate<chloride<bromide<nitrate<iodide. Low activities of guanidinium fluoride and sulphate were attributable to counter-effects of the anions, which acted as structural stabilizers. Changes in renaturation kinetics induced by either temperature or added perturbants appeared to conform with the Flory–Weaver model for the collagen transition. Additivity and non-specificity of the observed effects are discussed with particular reference to a common mechanism involving weak, non-saturated binding of perturbants at protein peptide groups.

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Sources of Collagen for Biomaterials in Skin Wound Healing

Bioengineering ◽

10.3390/bioengineering6030056 ◽

2019 ◽

Vol 6 (3) ◽

pp. 56 ◽

Cited By ~ 24

Author(s):

Evan Davison-Kotler ◽

William S. Marshall ◽

Elena García-Gareta

Keyword(s):

Collagen Type ◽

Recombinant Expression ◽

Collagen Type I ◽

Skin Wound ◽

Type I ◽

Expression Systems ◽

Skin Wound Healing ◽

Skin Collagen ◽

Human Collagen ◽

Collagen Type Vii

Collagen is the most frequently used protein in the fields of biomaterials and regenerative medicine. Within the skin, collagen type I and III are the most abundant, while collagen type VII is associated with pathologies of the dermal–epidermal junction. The focus of this review is mainly collagens I and III, with a brief overview of collagen VII. Currently, the majority of collagen is extracted from animal sources; however, animal-derived collagen has a number of shortcomings, including immunogenicity, batch-to-batch variation, and pathogenic contamination. Recombinant collagen is a potential solution to the aforementioned issues, although production of correctly post-translationally modified recombinant human collagen has not yet been performed at industrial scale. This review provides an overview of current collagen sources, associated shortcomings, and potential resolutions. Recombinant expression systems are discussed, as well as the issues associated with each method of expression.

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Sequence Alignment between vWF and Peptides Inhibiting the vWF-collagen Interaction Does not Result in the Identification of a Collagen-binding Site in vWF

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614077 ◽

2000 ◽

Vol 84 (10) ◽

pp. 621-625 ◽

Cited By ~ 3

Author(s):

R. M. van der Plas ◽

G. Vandecasteele ◽

S. Vauterin ◽

E. G. Huizinga ◽

J. J. Sixma ◽

...

Keyword(s):

Binding Site ◽

Collagen Type ◽

Collagen Type I ◽

Type I ◽

Binding Studies ◽

Collagen Types ◽

Collagen Binding ◽

Human Collagen ◽

Rat Tail ◽

Calf Skin

SummaryWe previously found that two peptides (N- and Q-peptide) selected by phage display for binding to an anti-vWF antibody, were able to inhibit vWF-binding to collagen (1). The sequence of those peptides could be aligned with the sequence in vWF at position 1129-1136 just outside the A3-domain. As the peptides represent an epitope or mimotope of vWF for binding to collagen we next wanted to study whether the alignment resulted in the identification of a new collagen binding site in vWF. We mutated the 1129-1136 VWTLPDQC sequence in vWF to VATAPAAC. Expressing this mutant vWF (7.8-vWF) in a fur-BHK cell line resulted in well processed 7.8-vWF containing a normal distribution of molecular weight multimers. However, binding studies of this mutant vWF to rat tail, human and calf skin collagens type I, to human collagen types III and VI, revealed no decrease in vWF-binding to any of these collagens. Thus, although the N-and Q-peptides did inhibit the vWF-collagen interaction, the resulting alignment with the vWF sequence did not identify a collagen binding site, pointing out that alignments (although with a high percentage of identity) do not always result in identification of binding epitopes. However, suprisingly removal of the A3-domain or changing the vWF sequence at position 1129-1136 resulted in an increase of vWF-binding to human collagen type VI and to rat tail collagen type I, implying that these changes result in a different conformation of vWF with an increased binding to these collagens as a consequence.

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Differential anion effects on thermal stability of collagen in the dispersed and aggregated states

Biochemical Journal ◽

10.1042/bj1370599 ◽

1974 ◽

Vol 137 (3) ◽

pp. 599-602 ◽

Cited By ~ 4

Author(s):

A. E. Russell

Keyword(s):

Thermal Stability ◽

Acid Solution ◽

Thermal Transition ◽

Dilute Solution ◽

Dilute Acid ◽

Dilute Acid Solution ◽

Skin Collagen ◽

Collagen Molecules ◽

Thermal Stability Of ◽

Calf Skin

The effects of KCNS and KI on thermal transition temperatures of calf skin collagen molecules in dilute acid solution and precipitated collagen fibrils from the same source were compared as a function of salt concentration and pH. The two salts produced qualitatively similar effects on each collagen form, but the response shown by single collagen molecules in dilute solution differed from that observed for molecular aggregates present in native-type fibrils.

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