Isodityrosine cross-linking mediates insolubilization of cell walls in Chlamydomonas.
Enzymatic removal of the cell wall induces vegetative Chlamydomonas reinhardtii cells to transcribe wall genes and synthesize new hydroxyproline-rich glycoproteins (HRGPs) related to the extensins found in higher plant cell walls. A cDNA expression library made from such induced cells was screened with antibodies to an oligopeptide containing the (SP)x repetitive domains found in Chlamydomonas wall proteins. One of the selected cDNAs encodes an (SP)x-rich polypeptide that also displays a repeated YGG motif. Ascorbate, a peroxidase inhibitor, and tyrosine derivatives were shown to inhibit insolubilization of both the vegetative and zygotic cell walls of Chlamydomonas, suggesting that oxidative cross-linking of tyrosines is occurring. Moreover, insolubilization of both walls was concomitant with a burst in H2O2 production and in extracellular peroxidase activity. Finally, both isodityrosine and dityrosine were found in hydrolysates of the insolubilized vegetative wall layer. We propose that the formation of tyrosine cross-links is essential to Chlamydomonas HRGP insolubilization.
Waffenschmidt
S
S
Institut für Biochemie, Universität zu Köln, Germany.
Woessner
J P
JP
Beer
K
K
Goodenough
U W
UW
D000595:Amino Acid Sequence; D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism; D001483:Base Sequence; D002473:Cell Wall / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D000939:Epitopes; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D008969:Molecular Sequence Data; D009195:Peroxidase / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D012995:Solubility; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000378:metabolism*
D003432:Cross-Linking Reagents; D000939:Epitopes; D006023:Glycoproteins; D012333:RNA, Messenger; D014443:Tyrosine; C035129:isodityrosine; D006861:Hydrogen Peroxide; C007543:dityrosine; D009195:Peroxidase; D001205:Ascorbic Acid