scholarly journals Effect of L-azetidine-2-carboxylic acid on glycosylations of collagen in chick-embryo tendon cells

1976 ◽  
Vol 160 (3) ◽  
pp. 639-645 ◽  
Author(s):  
A Oikarinen ◽  
H Anttinen ◽  
K I Kivirikko
Keyword(s):  
Carboxylic Acid ◽  
Chick Embryo ◽  
Type I Collagen ◽  
Type I ◽  
Pulse Label ◽  
Chase Period ◽  
Helix Formation ◽  
Collagen Secretion ◽  
Tendon Cells ◽  
Triple Helix Formation

The glycosylations of hydroxylysine during collagen biosynthesis in isolated chick-embryo tendon cells were studied by using pulse-chase labelling experiments with [14C]-lysine. The hydroxylation of lysine and the glycosylations of hydroxylysine continued after a 5 min pulse label for up to about 10 min during the chase period. These data differ from those obtained previously in isolated chick-embryo cartilage cells, in which, after a similar 5 min pulse label, these reactions continued during the chase period for up to about 20 min. The collagen synthesized by the isolated chick-embryo tendon cells differed markedly from the type I collagen of adult tissues in its degree of hydroxylation of lysine residues and glycosylations of hydroxylysine residues. When the isolated tendon cells were incubated in the presence of L-azetidine-2-carboxylic acid, the degree of glycosylations of hydroxylysine during the first 10 min of the chase period was identical with that in cells incubated without thcarboxylic acid for at least 60 min, whereas no additional glycosylations took place in the control cells after the 10 min time-point. As a consequence, the collagen synthesized in the presence of this compound contained more carbohydrate than did the collagen synthesized by the control cells. Additional experiments indicated that azetidine-2-carboxylic acid did not increase the collagen glycosyltransferase activities in the tendon cells or the rate of glycosylation reactions when added directly to the enzyme incubation mixture. Control experiments with colchicine indicated that the delay in the rate of collagen secretion, which was observed in the presence of azetidine-2-carboxylic acid, did not in itself affect the degree of glycosylations of collagen. The results thus suggest that the increased glycosylations were due to inhibition of the collagen triple-helix formation, which is known to occur in the presence of azetidine-2-carboxylic acid.

scholarly journals Effect of prevention of procollagen triple-helix formation on proline 3-hydroxylation in freshly isolated chick-embryo tendon cells

1981 ◽  
Vol 196 (1) ◽  
pp. 203-206 ◽  
Author(s):  
K Majamaa
Keyword(s):  
Carboxylic Acid ◽  
Chick Embryo ◽  
Triple Helix ◽  
Hydroxylase Activity ◽  
Intact Cells ◽  
Helix Formation ◽  
Tendon Cells ◽  
Triple Helix Formation

Inhibition of procollagen triple-helix formation by the addition of cis-hydroxyproline or azetidine-2-carboxylic acid increased the synthesis of 3-hydroxy[14C]proline 1.7-1.8-fold in pulse-chase experiments with freshly isolated chick-embryo tendon cells. The amount of 3-hydroxy[14C]proline, expressed as a percentage of the total 14C radioactivity in hydroxyproline, reached 8.4%. Control experiments indicated that the two analogues had no effect on the prolyl 3-hydroxylase activity of these cells. The data suggest that the time available before triple-helix formation in part regulates the extent of the 3-hydroxylation of proline in the biosynthesis of collagen in intact cells.

scholarly journals Embryonic Lethality of Molecular Chaperone Hsp47 Knockout Mice Is Associated with Defects in Collagen Biosynthesis

2000 ◽  
Vol 150 (6) ◽  
pp. 1499-1506 ◽  
Author(s):  
Naoko Nagai ◽  
Masanori Hosokawa ◽  
Shigeyoshi Itohara ◽  
Eijiro Adachi ◽  
Takatoshi Matsushita ◽  
...  
Keyword(s):  
Molecular Chaperone ◽  
Knockout Mice ◽  
Triple Helix ◽  
Type I Collagen ◽  
Molecular Form ◽  
Basement Membranes ◽  
Type I ◽  
Collagen Biosynthesis ◽  
Helix Formation ◽  
Triple Helix Formation

Triple helix formation of procollagen after the assembly of three α-chains at the C-propeptide is a prerequisite for refined structures such as fibers and meshworks. Hsp47 is an ER-resident stress inducible glycoprotein that specifically and transiently binds to newly synthesized procollagens. However, the real function of Hsp47 in collagen biosynthesis has not been elucidated in vitro or in vivo. Here, we describe the establishment of Hsp47 knockout mice that are severely deficient in the mature, propeptide-processed form of α1(I) collagen and fibril structures in mesenchymal tissues. The molecular form of type IV collagen was also affected, and basement membranes were discontinuously disrupted in the homozygotes. The homozygous mice did not survive beyond 11.5 days postcoitus (dpc), and displayed abnormally orientated epithelial tissues and ruptured blood vessels. When triple helix formation of type I collagen secreted from cultured cells was monitored by protease digestion, the collagens of Hsp47+/+ and Hsp47+/− cells were resistant, but those of Hsp47−/− cells were sensitive. These results indicate for the first time that type I collagen is unable to form a rigid triple-helical structure without the assistance of molecular chaperone Hsp47, and that mice require Hsp47 for normal development.

Delayed Triple-Helix Formation of Abnormal Type I Collagen Is Corrected by Reduced Temperature.

1988 ◽  
Vol 543 (1 Third Interna) ◽  
pp. 85-92 ◽  
Author(s):  
A. SUPERTI-FURGA ◽  
P. M. ROYCE ◽  
F. M. PISTONE ◽  
C. ROMANO ◽  
B. STEINMANN
Keyword(s):  
Triple Helix ◽  
Type I Collagen ◽  
Type I ◽  
Helix Formation ◽  
Triple Helix Formation ◽  
Reduced Temperature

scholarly journals Further studies on the effect of the collagen triple-helix formation on the hydroxylation of lysine and the glycosylations of hydroxylysine in chick-embryo tendon and cartilage cells

1977 ◽  
Vol 166 (3) ◽  
pp. 357-362 ◽  
Author(s):  
Aarne Oikarinen ◽  
Henrik Anttinen ◽  
Kari I. Kivirikko
Keyword(s):  
Chick Embryo ◽  
Triple Helix ◽  
Collagen Triple Helix ◽  
Helix Formation ◽  
Tendon Cells ◽  
Triple Helix Formation ◽  
Lysine Residues ◽  
Cartilage Cells ◽  
Helical Molecules ◽  
And Cartilage

The hydroxylation of lysine and glycosylations of hydroxylysine were studied in isolated chick-embryo tendon and cartilage cells under conditions in which collagen triple-helix formation was either inhibited or accelerated. The former situation was obtained by incubating the tendon cells with 0.6mm-dithiothreitol, thus decreasing their proline hydroxylase activity by about 99%. After labelling with [14C]proline, the formation of hydroxy[14C]proline was found to have declined by about 95%. Since the hydroxylation of a relatively large number of proline residues is required for triple-helix formation at 37°C, the pro-α-chains synthesized under these conditions apparently cannot form triple-helical molecules. Labelling experiments with [14C]lysine indicated that the degree of hydroxylation of the lysine residues in the collagen synthesized was slightly increased and the degree of the glycosylations of the hydroxylysine residues more than doubled, the largest increase being in the content of glucosylgalactosylhydroxylysine. Recovery of chick-embryo cartilage cells from temporary anoxia was used to obtain accelerated triple-helix formation. A marked decrease was found in the extent of hydroxylation of the lysine residues in the collagen synthesized under these conditions, and an even larger decrease occurred in the glycosylations of the hydroxylysine residues. The results support the previous suggestion that the triple-helix formation of the pro-α-chains prevents further hydroxylation of lysine residues and glycosylations of hydroxylysine residues during collagen biosynthesis.

HSP47, a collagen-specific chaperone protein, is a limiting factor for type I collagen secretion in aged skin

2017 ◽  
Vol 86 (2) ◽  
pp. e92
Author(s):  
MinJu Pyo ◽  
Jun Sang Park ◽  
Young Hun Lee ◽  
Dong Hun Lee ◽  
Jin Ho Chung ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Limiting Factor ◽  
Type I ◽  
Chaperone Protein ◽  
Collagen Secretion ◽  
Aged Skin

Changes in the transferrin requirement of cultured chick embryo mesoderm cells during early differentiation

Development ◽  
1986 ◽  
Vol 95 (1) ◽  
pp. 81-93
Author(s):  
E. J. Sanders
Keyword(s):  
Chick Embryo ◽  
Type I Collagen ◽  
Specific Effect ◽  
Primitive Streak ◽  
Cellular Adhesion ◽  
Type I ◽  
Surface Binding ◽  
Surface Receptors ◽  
Early Differentiation ◽  
Species Specific

Mesodermal tissue from the chick embryo at various stages of early differentiation was cultured in hydrated gels of type I collagen in the presence and absence of transferrin. Primary mesoderm explants from primitive-streak-stage embryos responded to the presence of avian transferrin by significantly improved outgrowth which appeared to be related to the ability of the cells to attach to, and migrate in, the collagen. No evidence was obtained which suggested that this observation was dependent on increased cell proliferation. This outgrowth enhancement was not duplicated by transferrin of human origin. The avian transferrin did not produce this effect on cells cultured on plastic substrata, suggesting that the species-specific effect involves modulation by the extracellular matrix. Mesoderm explants from somite stages of development showed no increase in outgrowth in the presence of either avian or human transferrin as judged by counting the number of outwandering cells. Ultrastructural immunocytochemistry indicated surface binding of transferrin by cells in the gels, and the presence of endogenous transferrin on the surfaces of mesoderm cells in situ and in their extracellular environment. It is suggested that by binding to cell surface receptors, transferrin may be able to influence the strength of cellular adhesion to collagen and hence the capacity for cell locomotion.

scholarly journals Superoxide-induced Type I collagen secretion depends on prolyl 4-hydroxylases

2020 ◽  
Vol 529 (4) ◽  
pp. 1011-1017
Author(s):  
Run Shi ◽  
Shanshan Gao ◽  
Andrew H. Smith ◽  
Huan Li ◽  
Ming Shao ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Collagen Secretion

scholarly journals Involvement of the stress protein HSP47 in procollagen processing in the endoplasmic reticulum

1992 ◽  
Vol 117 (4) ◽  
pp. 903-914 ◽  
Author(s):  
A Nakai ◽  
M Satoh ◽  
K Hirayoshi ◽  
K Nagata
Keyword(s):  
Endoplasmic Reticulum ◽  
Stress Protein ◽  
Normal Growth ◽  
Growth Conditions ◽  
Binding Activity ◽  
Pulse Label ◽  
Intact Cells ◽  
Collagen Binding ◽  
Chase Period ◽  
Helix Formation

The 47,000-D collagen-binding glycoprotein, heat shock protein 47 (HSP47), is a stress-inducible protein localized in the ER of collagen-secreting cells. The location and collagen-binding activity of this protein led to speculation that HSP47 might participate in collagen processing. Chemical crosslinking studies were used to test this hypothesis both before and after the perturbation of procollagen processing. The association of procollagen with HSP47 was demonstrated using cleavable bifunctional crosslinking reagents. HSP47 and procollagen were shown to be coprecipitated by the treatment of intact cells with anti-HSP47 or with anticollagen antibodies. Furthermore, several proteins residing in the ER were noted to be crosslinked to and coprecipitated with HSP47, suggesting that these ER-resident proteins may form a large complex in the ER. When cells were heat shocked, or when stable triple-helix formation was inhibited by treatment with alpha,alpha'-dipyridyl, coprecipitation of procollagen with HSP47 was increased. This increase was due to the inhibition of procollagen secretion and to the accumulation of procollagen in the ER. Pulse label and chase experiments revealed that coprecipitated procollagen was detectable as long as procollagen was present in the endoplasmic reticulum of alpha,alpha'-dipyridyl-treated cells. Under normal growth conditions, coprecipitated procollagen was observed to decrease after a chase period of 10-15 min, whereas total procollagen decreased only after 20-25 min. In addition, the intracellular association between HSP47 and procollagen was shown to be disrupted by a change in physiological pH, suggesting that the dissociation of procollagen from HSP47 is pH dependent. These findings support a specific role for HSP47 in the intracellular processing of procollagen, and provide evidence of a new category of "molecular chaperones" in terms of its substrate specificity and the dissociation mechanism.

The detrimental effect of orotic acid substitution in the peptide nucleic acid strand on the stability of PNA2:NA triple helices

2005 ◽  
Vol 83 (10) ◽  
pp. 1731-1740 ◽  
Author(s):  
Robert HE Hudson ◽  
Filip Wojciechowski
Keyword(s):  
Nucleic Acid ◽  
Carboxylic Acid ◽  
Peptide Nucleic Acid ◽  
Triple Helix ◽  
Orotic Acid ◽  
Uv Spectroscopy ◽  
Triplex Formation ◽  
Helix Formation ◽  
Triple Helix Formation ◽  
The Stability

We have investigated the incorporation of C6 derivatives of uracil into polypyrimidine peptide nucleic acid oligomers. Starting with uracil-6-carboxylic acid (orotic acid), a peptide nucleic acid monomer compatible with Fmoc-based synthesis was prepared. This monomer then served as a convertible nucleobase whereupon treatment of the resin-bound methyl orotate containing hexamers with hydroxide or amines cleanly converted the ester to an orotic acid or orotamide-containing peptide nucleic acid. Peptide nucleic acid hexamers containing the C6-modified nucleobase hybridized to both poly(riboadenylic acid) and poly(deoxyriboadenylic acid) via triplex formation. Complexes formed with poly(riboadenylic acid) were more stable than those formed with poly(dexoyriboadenylic acid), as measured by temperature-dependent UV spectroscopy. However, both of these complexes were destabilized relative to the complexes formed by an unmodified peptide nucleic acid oligomers. Internal or doubly substituted hexamers are destabilized more strongly than a terminally substituted one, and the type of substitution (carboxamide, ester, carboxylic acid) affects the overall triplex stability. These results clearly show that incorporation of a C6-substituted uracil into polypyrimidine PNA is detrimental to triplex formation. We have also extended this chemistry to incorporate uracil-5-methylcarboxylate into a peptide nucleic acid hexamer. After on-resin conversion of the C5 ester to the 3-(N,N-dimethylamino)propylamide, significant stabilization of the triplex formed with poly(riboadenylic acid) was observed, which illustrates the compatibility of C5 substitution with peptide nucleic acid directed triple helix formation. Key words: peptide nucleic acid, triple helix, orotic acid, orotamide, PNA.

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