Partial characterization of a collagen-binding, differentiation-related glycoprotein from skeletal myoblasts

Nandan, Devki; Cates, G A; Ball, Eric H.; Sanwal, B. D.

doi:10.1016/0003-9861(90)90263-x

Cited by 22 publications

(20 citation statements)

References 29 publications

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“…Hsp47 is a heat shock protein, present exclusively in the ER, that plays a vital role in procollagen processing. Hsp47 is expressed only by collagen-producing cells (8,9) and in vitro binds to collagens type I to V as well as gelatin (10). Within the ER, Hsp47 has been found to interact with nascent type I procollagen chains (11,12), with fully translated pro␣ collagen chains (13), with non-helical and poorly hydroxylated procollagen trimers (14) and with well-hydroxylated, triple helical procollagen (15,16).…”

mentioning

confidence: 99%

Functional Linkage between the Endoplasmic Reticulum Protein Hsp47 and Procollagen Expression in Human Vascular Smooth Muscle Cells

Rocnik¹,

Veer

Cao

et al. 2002

Journal of Biological Chemistry

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Hsp47 is a heat stress protein that interacts with procollagen in the lumen of the endoplasmic reticulum, which is vital for collagen elaboration and embryonic viability. The precise actions of Hsp47 remain unclear, however. To evaluate the effects of Hsp47 on collagen production we infected human vascular smooth muscle cells (SMCs) with a retrovirus containing Hsp47 cDNA. SMCs overexpressing Hsp47 secreted type I procollagen faster than SMCs transduced with empty vector, yielding a greater accumulation of pro␣1(I) collagen in the extracellular milieu. Interestingly, the amount of intracellular pro␣1(I) collagen was also increased. This was associated with an unexpected increase in the rate of pro␣1(I) collagen chain synthesis and 2.5-fold increase in pro␣1(I) collagen mRNA expression, without a change in fibronectin expression. This amplification of procollagen expression, synthesis, and secretion by Hsp47 imparted SMCs with an enhanced capacity to elaborate a fibrillar collagen network. The effects of Hsp47 were qualitatively distinct from, and independent of, those of ascorbate and the combination of both factors yielded an even more intricate fibril network. Given the in vitro impact of altered Hsp47 expression on procollagen production, we sought evidence for interindividual variability in Hsp47 expression and identified a common, single nucleotide polymorphism in the Hsp47 gene promoter among African Americans that significantly reduced promoter activity. Together, these findings indicate a novel means by which type I collagen production is regulated by the endoplasmic reticulum constituent, Hsp47, and suggest a potential basis for inherent differences in collagen production within the population.

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confidence: 99%

Functional Linkage between the Endoplasmic Reticulum Protein Hsp47 and Procollagen Expression in Human Vascular Smooth Muscle Cells

Rocnik¹,

Veer

Cao

et al. 2002

Journal of Biological Chemistry

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“…Antibodies against rat myoblast gp46 cross-react with colligin, but they do not react with the collagenbinding glycoprotein of Mr 47000 (hsp47) from chick-embryo fibroblasts (Nagata & Yamada, 1986). The published N-terminal sequences of rat myoblast gp46 and chick fibroblast hsp47 have no sequence similarity (Nagata et al, 1988;Nandan et al, 1990); however, binding properties and cellular location would suggest a similar function. The presence of gp46 or related proteins in the endoplasmic reticulum of collagen-secreting cells suggests that gp46 may play a role in intracellular assembly, transport, or secretion of collagen.…”

Section: Introductionmentioning

confidence: 99%

“…This glycoprotein is a heat-shock protein that represents approx. 1 % of the total cell protein in cultured myoblasts active in collagen synthesis (Nandan et al, 1990). Undifferentiated myoblasts synthesize gp46 coincident with type I collagen production, but, following differentiation into multinucleated myotubes, both synthesis of gp46 and collagen declines (Cates et al, 1987).…”

Section: Introductionmentioning

confidence: 99%

“…We have previously purified gp46 from rat myoblasts in a denatured form for a preliminary characterization (Nandan et al, 1990). In order to be able to examine the function of gp46, we have now purified gp46 in an active state which retains its collagen-binding property.…”

Section: Introductionmentioning

confidence: 99%

“…The endoplasmic reticulum of rat skeletal myoblasts as well as a number of other cell types contains a major collagen-binding glycoprotein of M, 46000 which has been designated gp46 (Cates et al, 1984(Cates et al, , 1987Nandan et al, 1988Nandan et al, , 1990). This glycoprotein is a heat-shock protein that represents approx.…”

Section: Introductionmentioning

confidence: 99%

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Purification and reconstitution of a collagen-binding heat-shock glycoprotein from L6 myoblasts

Vaillancourt

Cates

1991

Biochemical Journal

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A major collagen-binding heat-shock glycoprotein from L6 myoblasts, designated gp46, was purified by gelatin-agarose chromatography and ion-exchange chromatography. Purified gp46 was functionally active, as shown by its ability to rebind to gelatin-agarose, and was homogeneous as determined by SDS/polyacrylamide-gel electrophoresis. This is the first reported purification of myoblast gp46 in an active state. Triton X-100-soluble gp46 was found to bind preferentially to immobilized pepsin-treated type IV collagen compared with native type I collagen. gp46, reconstituted into phospholipid vesicles, retained collagen-binding activity. This activity could be destroyed by chemical modification with chloramine-T, but was decreased by only 20(30 % following treatment with iodoacetamide or N-ethylmaleimide. Since gp46 is a heat-shock protein, we examined the possibility that it may confer protection on type I collagen from thermal denaturation at temperatures above its normal melting temperature of 41 'C. In the presence of gp46 liposomes the apparent melting temperature of type I collagen was marginally increased to 42 'C. This change was considered to be too small to be of physiological significance. We have therefore concluded that the role of gp46 in collagen metabolism is unlikely to be related to any thermal-stabilizing function.

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Non‐natural CBP2 binding peptides and peptomers modulate carcinoma cell adhesion and invasion

Hebert

Coletta

Norris

et al. 2001

J of Cellular Biochemistry

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A combinatorial approach that utilized a repertoire of bacteriophage-peptides has identified a number of non-natural CBP2 binding peptides. Moreover, co-localization of some of these peptides with CBP2 in a number of tumor cell lines demonstrated that the peptides were directed to an intracellular location spatially coincident with the normal distribution of CBP2 [Sauk et al., 2000]. From among these sequences WHYPWFQNWAMA and LDSRYSLQAAMY were the most effective CBP2 binding peptides and best fulfilled the combinatorial motif containing deep hydrophobic pockets. When the hydropathic profiles of collagen alpha1(IV) and alpha2 (IV) were compared with these dodecapeptides, the hydropathic profiles of WHYPWFQNWAMA and LDSRYSLQAAMY closely matched those of alpha1(IV) 414-452 and alpha1(IV)531-543. These peptides were shown to be functional peptidomimics and possessed the ability to alter cell adhesion and invasion of human squamous cell carcinoma cell lines. Peptomers were formed of these non-natural peptides to explore the role that a repetitive peptide may have on cell adhesion. The enhanced cell adhesion observed with the peptomers required both CBP2 antibodies and integrin antibodies for inhibition. The enhanced adhesion observed even in the face of combined antibody inhibition was consistent with such complexes possessing correspondingly slower dissociation rates. Thus, suggesting that peptomers may function in a like manner to multimeric peptide MHC complexes (tetramers) binding more than one cell receptor on a specific cell. These findings evoke both peptidomimics of native ligands and their peptomers as potential reagents by which to target tumor cells for chemotherapy, imaging, or retargeting viral vectors for gene therapy.

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Partial characterization of a collagen-binding, differentiation-related glycoprotein from skeletal myoblasts

Cited by 22 publications

References 29 publications

Functional Linkage between the Endoplasmic Reticulum Protein Hsp47 and Procollagen Expression in Human Vascular Smooth Muscle Cells

Functional Linkage between the Endoplasmic Reticulum Protein Hsp47 and Procollagen Expression in Human Vascular Smooth Muscle Cells

Purification and reconstitution of a collagen-binding heat-shock glycoprotein from L6 myoblasts

Non‐natural CBP2 binding peptides and peptomers modulate carcinoma cell adhesion and invasion

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