Properties of radiolabeled type I, II, and III collagens related to their use as substrates in collagenase assays

Journal Article · Sat Nov 01 00:00:00 EST 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90557-9· OSTI ID:6943008

Mookhtiar, K A; Mallya, S K; Van Wart, H E

Calf skin and rat tendon type I, bovine cartilage type II, and human amnion type III collagens have been radiolabeled by reaction with (/sup 3/H)acetic anhydride, (/sup 3/H)formaldehyde, and succinimidyl 2,3-(3H)propionate. All three reactions produce collagens with high specific activities that are suitable for use as substrates in collagenase assays. The identity of the radiolabel and the labeling indices do not alter the molecular weights or thermal stabilities of the collagens or the solubilities of the collagens or gelatins in dioxane-water mixtures at 4 degrees C. However, in contrast to native or sparsely labeled collagens, those with 40 or more lysine + hydroxylysine residues labeled per molecule do not undergo fibrillogenesis in the presence of 0.2-0.4 M NaCl in the 4-35 degree C temperature range. Thus, the modification reactions not only serve to introduce the radiolabel, but also to keep the collagens soluble over a wide range of temperatures and concentrations. The TCA, TCB fragments produced on partial reaction of each collagen type with tissue collagenases can be selectively denatured by a 10-minute incubation under specific conditions and the intact collagens selectively precipitated by addition of 50% v/v dioxane. This serves as the basis for soluble collagenase assays. The effect of labeling index on the properties of the collagens has been investigated and the results establish the range of conditions over which these collagens can be used as substrates for soluble versus fibrillar collagenase assays.

• Research Organization: Florida State Univ., Tallahassee
• OSTI ID: 6943008
• Journal Information: Anal. Biochem.; (United States), Vol. 2
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PEPTIDE HYDROLASES
LABELLING
ACETYLATION
ACYLATION
CATTLE
COLLAGEN
GELS
RADIOBIOLOGY
RADIOENZYMATIC ASSAY
RATS
SOLUBILITY
SUBSTRATES
TRITIUM COMPOUNDS
ANIMALS
BIOLOGY
CHEMICAL REACTIONS
COLLOIDS
DISPERSIONS
DOMESTIC ANIMALS
ENZYMES
HYDROLASES
LABELLED COMPOUNDS
MAMMALS
ORGANIC COMPOUNDS
PROTEINS
RODENTS
RUMINANTS
SCLEROPROTEINS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques