Characterization of three constituent chains of collagen type VI by peptide sequences and cDNA clones.
Pepsin-solubilized collagen VI was prepared from human placenta and used to separate three constituent chains for determining partial amino acid sequences. Antibodies raised against the chains assisted in the identification and purification of several cDNA clones from three expression lambda gt11 libraries. Most of the clones hybridized to either a 3.5-kb or 4.2-kb mRNA species which by matching peptide and nucleotide sequences could be identified as coding for the alpha 2(VI) or alpha 1(VI) chain, respectively. Other clones hybridized to either an 8.5-kb mRNA which very likely encoded the alpha 3(VI) chain or to an unknown 2.0-kb mRNA. Northern blots revealed a considerable variation in the mRNA levels for each collagen VI chain in both skin and cornea fibroblasts and in several tumor cell lines. Limited sequence data generated from peptides and cDNA clones demonstrated a characteristic cysteine pattern at the junction between N-terminal globular domain and triple helix in all three chains. In addition, the data showed occasional interruptions of triplet sequences within the triple-helical domain and the presence of two Arg-Gly-Asp sequences which are potential cell-binding structures.
Chu
M L
ML
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania.
Mann
K
K
Deutzmann
R
R
Pribula-Conway
D
D
Hsu-Chen
C C
CC
Bernard
M P
MP
Timpl
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics / Q000302:isolation & purification*; D004247:DNA / Q000032:analysis*; D004586:Electrophoresis; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010449:Peptide Mapping; D010454:Peptide Termination Factors / Q000032:analysis / Q000235:genetics; D010920:Placenta / Q000032:analysis; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits
D010454:Peptide Termination Factors; D012333:RNA, Messenger; D003094:Collagen; D004247:DNA