Structural study of long arm fragments of laminin. Evidence for repetitive C-terminal sequences in the A-chain, not present in the B-chains.
The outer segments of the long arm of laminin have recently been shown to mediate attachment of many cell types and to stimulate neurite outgrowth. For a structural characterization of this part of the molecule we prepared, by limited elastase digestion of laminin, fragments E3 and E8, previously identified as a globular heparin-binding domain and as a 35-nm-long rod with a terminal globule, respectively. Fragment E3 is a domain adjacent to fragment E8. Both structures together comprise the complete terminal half of the long arm. Our data confirm current models, which predict that the C-terminal segments from all three chains contribute to its structure. The B chains terminate at the end of the rod like domain, while the large terminal globule is formed by A-chain structures only. In addition to fragment E3, two new fragments T1 and T2 obtained by tryptic cleavage of fragment E8 were characterized as substructures of the globular domain. Screening of a mouse cDNA library with synthetic oligonucleotides allowed isolation of an 1.8-kb cDNA clone encoding 547 C-terminal amino acids of the A chain and some 196 nucleotides of the 3'-untranslated region including a single polyadenylation site. The clone contained portions of domain T2 and the complete heparin binding domain E3 which was thus identified as the most C-terminal domain of the A chain. Sequence alignment indicated that the terminal globule is formed by homologous repeats of some 140 residues having no counterpart in the B chains.
Deutzmann
R
R
Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
Huber
J
J
Schmetz
K A
KA
Oberbäumer
I
I
Hartl
L
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002845:Chromatography / Q000379:methods; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D004586:Electrophoresis / Q000379:methods; D007797:Laminin / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010196:Pancreatic Elastase; D012333:RNA, Messenger / Q000032:analysis; D012689:Sequence Homology, Nucleic Acid
D007797:Laminin; D012333:RNA, Messenger; D004247:DNA; D010196:Pancreatic Elastase