Skip to main content
PMC home page
Cytotechnology logoLink to Cytotechnology
. 2000 Jul;33(1-3):167–174. doi: 10.1023/A:1008186912975

Three heterotrimeric laminins produced by human keratinocytes

Chino Kumagai 1, Masaki Okano 3, Yasuo Kitagawa 1,✉
PMCID: PMC3466727  PMID: 19002824

Abstract

Laminins are a family of glycoproteins composed of α,β and γ chains. Five α(α1-α5), three β (β1–β3) and twoγ (γ1 and γ2) chains have been cloned fromhuman and their replaceable assembly into heterotrimers producesthe variety of laminins. Reverse transcription-polymerase chainreaction of mRNAs showed that human keratinocytes express theα3, α5, β1, β3, γ1 andγ2 genes at high level among the ten cloned lamininchains. Western blot and immunoprecipitation of the cell lysatewith antiserum directed against mouse laminin-1(α1β1γ1) detected two trimers with thecomposition of αxβ1γ1 (probablylaminin-10 with the composition of α5β1γ1and αyβ1γ1. Meanwhile, antiserum directedagainst a synthetic peptide of human α3 detected onlyα3β3γ2 trimer (laminin-5). We thus show thatkeratinocytes produce three heterotrimeric laminins. We couldnot detect the assembly of α3 with β1 and γ1chains to form α3β1γ1 (laminin-6) in keratinocytes.

Keywords: epiligrin, kalinin, keratinocyte, laminin

Full Text

The Full Text of this article is available as a PDF (92.7 KB).

References

  1. Aratani Y, Kitagawa Y. Enhanced synthesis and secretion of type IV collagen and entactin during adipose conversion of 3T3-L1 cells and production of unorthodox laminin complex. J Biol Chem. 1988;263:16163–16169. [PubMed] [Google Scholar]
  2. Beck K, Hunter I, Engel J. Structure and function of laminin: Anatomy of a multidomain glycoprotein. FASEB J. 1990;4:148–160. doi: 10.1096/fasebj.4.2.2404817. [DOI] [PubMed] [Google Scholar]
  3. Burgeson RE, Chiquet M, Deutzmann R, Ekblom P, Engel J, Kleinman H, Martin GR, Meneguzzi G, Paulsson M, Sanes J, Timpl R, Tryggvason K, Yamada Y, Yurchenco PD. A new nomenclature for the laminins. Matrix Biol. 1994;14:209–211. doi: 10.1016/0945-053x(94)90184-8. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Durkin ME, Loechel F, Mattei MG, Gilpin BJ, Albrechtsen R, Wewer UM. Tissue-specific expression of the human laminin α5-chain, and mapping of the gene to distal mouse chromosome 2 near the locus for the ragged (Ra) mutation. FEBS Lett. 1997;411:296–300. doi: 10.1016/s0014-5793(97)00686-8. [DOI] [PubMed] [Google Scholar]
  6. Engvall E, Wewer UM. Domains of laminin. J Cell Biochem. 1996;61:493–501. doi: 10.1002/(SICI)1097-4644(19960616)61:4%3C493::AID-JCB2%3E3.0.CO;2-J. [DOI] [PubMed] [Google Scholar]
  7. Gerecke DR, Wagman DW, Champliaud MF, Burgeson RE. The complete primary structure for a novel laminin chain, the laminin B1k chain. J Biol Chem. 1994;269:11073–11080. [PubMed] [Google Scholar]
  8. Iivanainen A, Vuolteenaho R, Sainio K, Eddy R, Shows TB, Sariola H, Tryggvason K. The human laminin β2 chain (Slaminin): Structure, expression in fetal tissues and chromosomal assignment of the LAMB2 gene. Matrix Biol. 1995;14:489–497. doi: 10.1016/0945-053x(95)90006-3. [DOI] [PubMed] [Google Scholar]
  9. Kallunki P, Sainio K, Eddy R, Byers M, Kallunki T, Sariola H, Beck K, Hirvonen H, Shows TB, Tryggvason KA. Truncated laminin chain homologous to the B2 chain: Structure, spatial expression, and chromosomal assignment. J Cell Biol. 1992;119:679–693. doi: 10.1083/jcb.119.3.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Liu J, Mayne R. The complete cDNA coding sequence and tissue-specific expression of the mouse laminin α4 chain. Matrix Biol. 1996;15:433–437. doi: 10.1016/s0945-053x(96)90162-6. [DOI] [PubMed] [Google Scholar]
  11. Marinkovich MP, Lunstrum GP, Keene DR, Burgeson RE. The dermal-epidermal junction of human skin contains a novel laminin variant. J Cell Biol. 1992;119:695–703. doi: 10.1083/jcb.119.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marinkovich MP, Lunstrum GP, Burgeson RE. The anchoring filament protein kalinin is synthesized and secreted as a high molecular weight precursor. J Biol Chem. 1992;267:17900–17906. [PubMed] [Google Scholar]
  13. Miki K, Sugimoto E, Kitagawa Y. Reversible interconversion between primitive endoderm-and partial endoderm-like F9 cells demonstrated by mRNAs expression. J Biochem (Tokyo) 1987;102:385–392. doi: 10.1093/oxfordjournals.jbchem.a122065. [DOI] [PubMed] [Google Scholar]
  14. Miner JH, Lewis RM, Sanes JR. Molecular cloning of a novel laminin, α5, and widespread expression in adult tissues. J Biol Chem. 1995;270:28523–28526. doi: 10.1074/jbc.270.48.28523. [DOI] [PubMed] [Google Scholar]
  15. Miner JH, Patton BL, Lentz SI, Gilbert DJ, Snider WD, Jenkins NA, Copeland NG, Sanes JR. The laminin α chains: Expression, developmental transitions, and chromosomal locations of α1-5, identification of heterotrimeric laminin 8-11, and cloning of a novel α3 isoform. J Cell Biol. 1997;137:685–701. doi: 10.1083/jcb.137.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morita A, Sugimoto E, Kitagawa Y. Post-translational assembly and glycosylation of laminin subunits in parietal endoderm-like F9 cells. Biochem J. 1985;229:259–264. doi: 10.1042/bj2290259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Niimi T, Miki K, Kitagawa Y. Expression of long arm sequence of mouse laminin α1, β1, or γ 1 chain in COS1 cells and assembly of monkey-mouse hybrid laminin. J Biochem (Tokyo) 1997;121:854–861. doi: 10.1093/oxfordjournals.jbchem.a021665. [DOI] [PubMed] [Google Scholar]
  18. Nissinen M, Vuolteenaho R, Boot-Handford R, Kallunki P, Tryggvason K. Primary structure of the human laminin A chain: Limited expression in human tissues. Biochem J. 1991;276:369–379. doi: 10.1042/bj2760369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nomura K, Sugawara T, Sato T, Sawamura D, Hashimoto I, Sugita Y, Uitto J. Expression of laminin, type IV procollagen and 230 kDa bullous pemphigoid antigen genes by keratinocytes and fibroblasts in culture: Application of the polymerase chain reaction for detection of small amounts of messenger RNA. Arch Dermatol Res. 1994;286:408–413. doi: 10.1007/BF00371801. [DOI] [PubMed] [Google Scholar]
  20. Pikkarainen T, Eddy R, Fukushima Y, Byers M, Shows T, Pihlajaniemi T, Saraste M, Tryggvason K. Human laminin B1 chain: A multidomain protein with gene (LAMB1) locus in the q22 region of chromosome 7. J Biol Chem. 1987;262:10454–10462. [PubMed] [Google Scholar]
  21. Pikkarainen T, Kallunki T, Tryggvason K. Human laminin B2 chain: Comparison of the complete amino acid sequence with the B1 chain reveals variety in sequence homology between different structural domains. J Biol Chem. 1988;263:6751–6758. [PubMed] [Google Scholar]
  22. Rousselle P, Lunstrum GP, Keene DR, Burgeson RE. Kalinin: An epithelium-specific basement membrane adhesion molecule that is a component of anchoring filaments. J Cell Biol. 1991;114:567–576. doi: 10.1083/jcb.114.3.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ryan MC, Tizard R, VanDevanter DR, Carter WD. Cloning of the LamA3 gene encoding the α3 chain of the adhesive ligand epiligrin. Expression in wound repair. J Biol Chem. 1994;269:22779–22787. [PubMed] [Google Scholar]
  24. Richards AJ, Al-Imara L, Carter NP, Lloyd JC, Leversha MA, Pope FM. Localization of the gene (LAMA4) to chromosome 6q21 and isolation of a partial cDNA encoding a variant laminin A chain. Genomics. 1994;22:237–239. doi: 10.1006/geno.1994.1372. [DOI] [PubMed] [Google Scholar]
  25. Timpl R, Brown JC. The laminins. Matrix Biol. 1994;14:275–281. doi: 10.1016/0945-053x(94)90192-9. [DOI] [PubMed] [Google Scholar]
  26. Tokida Y, Aratani Y, Morita A, Kitagawa T. Production of two variant laminin forms by endothelial cells and shift of their relative levels by angiostatic steroids. J Biol Chem. 1990;265:18123–18129. [PubMed] [Google Scholar]
  27. Vuolteenaho R, Nissinen M, Sainio K, Byers M, Eddy R, Hirvonen H, Shows TB, Sariola H, Engvall E, Tryggvason K. Human laminin M chain (merosin): Complete primary structure, chromosomal assignment, and expression of the M and A chain in human fetal tissues. J Cell Biol. 1994;124:381–394. doi: 10.1083/jcb.124.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES