Biochemical and Biophysical Research Communications
Identification and recombinant production of human laminin α4 subunit splice variants
Section snippets
Materials and methods
Cells and cell culture. The following cell lines were obtained from American Type Culture Collection (Manassas, VA): SK-N-SH and NB-1 (neuroblastoma); T98G, U251MG, U251SP, and AJ (glioma); HepG2 (hepatoblastoma); JEG3 (choriocarcinoma); A549 (lung carcinoma); 293 (embryonic kidney epithelial cells); and HT1080 (fibrosarcoma). All cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Sigma) supplemented with 10% fetal bovine serum (FBS) under an atmosphere of 95% air/5% CO2 and
Identification of a splice variant of human laminin α4 cDNA
A fragment of laminin α4 subunit cDNA with 21 extra bases was identified by sequencing the RT-PCR products amplified from the T98G glioma cells (Fig. 1A). This 21-bp insertion results in an in-frame insertion of seven amino acid residues, MDCPTIS, between Gly265 and Cys266 (amino acids are numbered according to [5]). Interestingly, a cysteine residue is present within the seven amino acid insert which is located very close to two other cysteines, Cys266 and Cys269. Since these cysteine residues
References (23)
The laminin family
Curr. Opin. Cell Biol.
(1993)- et al.
A new nomenclature for the laminins
Matrix Biol.
(1994) - et al.
Differential expression of laminin α chains during proliferative and differentiation stages in a model for skin morphogenesis
Matrix Biol.
(2000) - et al.
Primary structure and expression of a novel human laminin α4 chain
FEBS Lett.
(1995) - et al.
Blood platelets contain and secrete laminin-8 (α4β1γ1) and adhere to laminin-8 via α6β1 integrin
Exp. Cell Res.
(1999) - et al.
A splicing variant of steroid receptor coactivator-1 (SRC-1E): the major isoform of SRC-1 to mediate thyroid hormone action
Biochem. Biophys. Res. Commun.
(1997) - et al.
Purification and characterization of human laminin-8. Laminin-8 stimulates cell adhesion and migration through α3β1 and α6β1 integrins
J. Biol. Chem.
(2001) - et al.
Modification of the laminin α4 chain by chondroitin sulfate attachment to its N-terminal domain
FEBS Lett.
(2001) - et al.
Structure and function of laminin LG modules
Matrix Biol.
(2000) - et al.
Contributions of the LG modules and furin processing to laminin-2 functions
J. Biol. Chem.
(2002)
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Laminin isoform profiles in salivary glands in Sjögren's syndrome
2011, Advances in Clinical ChemistryCitation Excerpt :Cytokines and other molecules regulating LM transcription bind to enhancer or silencer elements found in promoters of LM genes, in the 5′ untranslated region, or inside introns as is the situation with murine LAMC1[57–60]. Additional dimension to LM regulation comes from alternative splicing, which has been demonstrated in almost all LM chains resulting in multiple isoforms of these chains [61–64]. Further, promoter methylation of LAMA3, LAMB3 and LAMC2 genes has been shown to decrease the expression of these genes in prostate and breast cancer [65,66].
Identification of a novel family of laminin N-terminal alternate splice isoforms: Structural and functional characterization
2009, Journal of Biological ChemistryCitation Excerpt :In contrast, LAMA3B contains exons 1–38 and 40–76 (i.e. skips exon 39), and encodes a full-length laminin termed α3b (Fig. 1B). Further alternate or minor isoforms have been identified in LAMC2 (18, 19), LAMA4 (20), LAMB3 (21), and LAMA2 (22). Here, we describe the identification of multiple, short, alternate splice isoforms derived from the 5′-end of the genes (LAMA3 and LAMA5) encoding the α3 and α5 laminin subunits.
The C-terminal region of laminin β chains modulates the integrin binding affinities of laminins
2009, Journal of Biological ChemistryLaminin isoforms containing the γ3 chain are unable to bind to integrins due to the absence of the glutamic acid residue conserved in the C-terminal regions of the γ1 and γ2 chains
2008, Journal of Biological ChemistryCitation Excerpt :Construction of Expression Vectors—Soluble clasped α6, α7X2, and β1 integrin expression vectors were prepared as described previously (10, 33). Expression vectors for the human laminin α1, α3, α4, α5, β1, β3, and γ1 chains were constructed as described (16, 20, 33, 34). Expression vectors for the human laminin α2 (GenBank™ accession number NM_000426), β2 (GenBank™ accession number NM_002292), and γ3 (GenBank™ accession number NM_006059) chains were prepared as follows.
Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells
2008, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Preparation of culture vessels coated with recombinant laminins. Five recombinant human laminin isoforms, rhLM-111, -211, -332, -411, and -511 were produced using the Free Style™ 293 Expression System (Invitrogen) and purified from conditioned medium as described previously [14,15]. The rhLMs were filtered through the 0.45-μm pores of Ultrafree-CL Centrifugal Filter Units (MILLIPORE) for sterile usage.
The requirement of the glutamic acid residue at the third position from the carboxyl termini of the laminin γ chains in integrin binding by laminins
2007, Journal of Biological ChemistryCitation Excerpt :αV and β3 integrin expression vectors were generously provided by Dr. Junichi Takagi (31, 32). Expression vectors for human laminin α5 chain lacking LG4-5 modules (pcDNA-α5ΔLG4-5), LG3–5 modules (pcDNA-α5ΔLG3–5), β1 chain (pCEP-β1), and γ1 chain (pcDNA3.1-γ1) were constructed as described (19, 33). Expression vectors for the laminin γ1 chain lacking the C-terminal 8 amino acids (γ1Δ8AA), 3 amino acids (γ1Δ3AA), 2 amino acids (γ1Δ2AA), or 1 amino acid (γ1Δ1AA) and that encoding the mutant γ1 chain with substitution of Gln for Glu-1607 (γ1EQ) were prepared as follows.