Identification of Cell Binding Sequences in Mouse Laminin γ1 Chain by Systematic Peptide Screening

Nomizu, Motoyoshi; Kuratomi, Yuichiro; Song, Sang‐Yong; Ponce, M. Lourdes; Hoffman, Matthew P.; Powell, Stephen Joseph; Miyoshi, Kengo; Otaka, Akira; Kleinman, Hynda K.; Yamada, Yoshihiko

doi:10.1074/jbc.272.51.32198

Cited by 109 publications

(128 citation statements)

References 43 publications

Supporting

Mentioning

103

Contrasting

Unclassified

Order By: Relevance

“…So far, five ␣, three ␤, and three ␥ chains have been identified, and at least 15 isoforms (laminin [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] are formed by various combinations of each subunit (1)(2)(3)(4). Laminins have diverse biological activities including promotion of cell adhesion, migration, neurite outgrowth, angiogenesis, and tumor metastasis (5).…”

mentioning

confidence: 99%

Identification of Biologically Active Sequences in the Laminin α4 Chain G Domain

Okazaki

Suzuki

Nishi

et al. 2002

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Laminins are a family of trimeric extracellular matrix proteins consisting of ␣, ␤, and ␥ chains. So far five different laminin ␣ chains have been identified. The laminin ␣4 chain, which is present in laminin-8/9, is expressed in cells of mesenchymal origin, such as endothelial cells and adipocytes. Previously, we identified heparin-binding sites in the C-terminal globular domain (G domain) of the laminin ␣4 chain. Here we have focused on the biological functions of the laminin ␣4 chain G domain and screened active sites using a recombinant protein and synthetic peptides. The rec-␣4G protein, comprising the entire G domain, promoted cell attachment activity. The cell attachment activity of rec-␣4G was completely blocked by heparin and partially inhibited by EDTA. We synthesized 116 overlapping peptides covering the entire G domain and tested their cell attachment activity. Twenty peptides showed cell attachment activity, and 16 bound to heparin. We further tested the effect of the 20 active peptides in competition assays for cell attachment and heparin binding to rec-␣4G protein. A4G6 (LAIKNDNLVYVY), A4G20 (DVIS-LYNFKHIY), A4G82 (TLFLAHGRLVFM), and A4G83 (LVFMFNVGHKKL), which promoted cell attachment and heparin binding, significantly inhibited both cell attachment and heparin binding to rec-␣4G. These results suggest that the four active sites are involved in the biological functions of the laminin ␣4 chain G domain. Furthermore, rec-␣4G, A4G6, and A4G20 were found to interact with syndecan-4. These active peptides may be useful for defining of the molecular mechanism laminin-receptor interactions and laminin-mediated cellular signaling pathways.Laminins, a family of extracellular matrix proteins, consist of three different subunits, ␣, ␤, and ␥ chains. So far, five ␣, three ␤, and three ␥ chains have been identified, and at least 15 isoforms (laminin 1-15) are formed by various combinations of each subunit (1-4). Laminins have diverse biological activities including promotion of cell adhesion, migration, neurite outgrowth, angiogenesis, and tumor metastasis (5). More than 20 receptors have been reported for these laminin molecules (6). Several active sites on laminin-1 have been identified using proteolytic fragments, recombinant proteins, and synthetic peptides (7,8). Previously, we screened for cell adhesive sequences on laminin-1 using 673 overlapping synthetic peptides covering the entire protein (9 -12). Most of the active peptides were localized in the globular domains and found to play a critical role in binding to cell surface receptors in a peptide-and cell type-specific manner (13,14). Several peptides were found to interact with integrins and syndecans (15-18). Some of the peptides promoted neurite outgrowth, angiogenesis, and tumor metastasis (19 -22).The laminin ␣ chains are generally large (M r ϭ 400,000) and contain a C-terminal globular domain consisting of five globular modules LG1-LG5. The laminin ␣4 chain lacks the N-terminal short arm and is expressed in cells of mesenchymal origin, such as...

show abstract

mentioning

confidence: 99%

Identification of Biologically Active Sequences in the Laminin α4 Chain G Domain

Okazaki

Suzuki

Nishi

et al. 2002

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Nine peptides active for cell adhesion from the mouse laminin b1 chain and seven peptides from the g1 chain (Nomizu et al, 1997) were tested for their effects on in vivo experimental pulmonary metastases of B16 -F10 mouse melanoma cells (Table 1). In control experiments (without peptides), the average of 163 metastatic colonies was formed on the surface of the lung within 16 days after B16 -F10 cells were injected into the mouse tail vein (Table 2).…”

Section: Resultsmentioning

confidence: 99%

“…Five peptides from the G-domain of a1 chain (Nomizu et al, 1995), 12 peptides from the g1 chain (Nomizu et al, 1997), 21 peptides from the short and long arms of a1 chain , and 12 peptides from the b1 chain were found to have significant cell adhesive activity. In addition, several peptides from the laminin-1 have been identified to be active for angiogenesis Ponce et al, 1999), acinar formation of salivary gland , and neurite outgrowth .…”

mentioning

confidence: 99%

Laminin γ1 chain peptide, C-16 (KAFDITYVRLKF), promotes migration, MMP-9 secretion, and pulmonary metastasis of B16–F10 mouse melanoma cells

et al. 2002

View full text Add to dashboard Cite

Laminin-1, a heterotrimer of a1, b1, and g1 chains specific to basement membrane, promotes cell adhesion and migration, proteinase secretion and metastases of tumour cells. Several active sites on the a1 chain have been found to promote B16 -F10 melanoma lung colonisation and here we have determined whether additional tumour promoting sites exist on the b1 and g1 chains. Recently, we have identified novel cell adhesive peptides derived from laminin b1 and g1 chains by systematic screening of synthetic peptides. Nine b1 peptides and seven g1 peptides active for cell adhesion were tested for their effects on experimental pulmonary metastases of B16 -F10 mouse melanoma cells in vivo. The most active adhesive peptide derived from the g1 chain globular domain, C-16 (KAFDITYVRLKF), significantly enhanced pulmonary metastases of B16 -F10 cells, whereas no other peptides showed enhancement. C-16 also stimulated migration of B16 -F10 cells in the Boyden chamber assay in vitro. Furthermore, C-16 significantly induced the production of MMP-9 from B16 -F10 cells. These results suggest that this specific laminin g1 chain peptide has a metastasis-promoting activity and might be a new molecular target of anti-cancer treatment.

show abstract

“…These multiple and diverse cell surface receptors bind to the laminins in different ways depending on the laminin isoforms. Various receptor-specific binding sequences in laminins have been identified in studies using recombinant proteins and synthetic peptides [11,[30][31][32][33][34][35][36]. The regulation of these laminin-receptor interactions is important for many biological processes.…”

Section: Introductionmentioning

confidence: 99%

“…For example, we previously identified about sixty cell-adhesive sequences from laminin-111 using 673 synthetic peptides covering the entire sequence. These active peptides promote various biological activities, including the promotion of cell spreading, cell differentiation, neurite outgrowth, angiogenesis and wound healing, and many of them bind to distinct cellular receptors [32][33][34][35]37,38].…”

Section: Introductionmentioning

confidence: 99%

Active Peptide-Conjugated Chitosan Matrices as an Artificial Basement Membrane

et al. 2015

Self Cite

View full text Add to dashboard Cite

Abstract:The basement membrane, a thin extracellular matrix, plays a critical role in tissue development and repair. Laminins are the major component of basement membrane and have diverse biological activities. We have identified various cell-adhesive peptides from laminins and their specific cell surface receptors. Polysaccharides, including chitosan, have been used as scaffolds, which regulate cellular functions for tissue engineering. We have developed laminin-derived active peptide-chitosan matrices as functional scaffolds. The biological activity of the peptides was enhanced when the peptides were conjugated to a chitosan matrix, suggesting that the peptide-chitosan matrix approach has an advantage for an active biomaterial. Further, the laminin peptide-chitosan matrices have the potential to mimic the basement membrane and are useful for tissue engineering as an artificial basement membrane.

show abstract

Identification of Cell Binding Sequences in Mouse Laminin γ1 Chain by Systematic Peptide Screening

Cited by 109 publications

References 43 publications

Identification of Biologically Active Sequences in the Laminin α4 Chain G Domain

Identification of Biologically Active Sequences in the Laminin α4 Chain G Domain

Laminin γ1 chain peptide, C-16 (KAFDITYVRLKF), promotes migration, MMP-9 secretion, and pulmonary metastasis of B16–F10 mouse melanoma cells

Active Peptide-Conjugated Chitosan Matrices as an Artificial Basement Membrane

Contact Info

Product

Resources

About