Construction and characterization of the soybean leaf metalloproteinase cDNA.
The cloning and analysis of a cDNA clone encoding the soybean metalloproteinase obtained by polymerase chain reaction (PCR) and the rapid amplification of cDNA ends (RACE) reaction are described. The cDNA was constructed from poly(A)+ RNA isolated from 15-17 day old leaves. The deduced amino acid sequence of the cDNA reveals that the plant metalloproteinase is synthesized as a preproenzyme and the proenzyme form shares a structural motif, responsible for maintenance of inactive zymogen, with the matrix metalloproteinase (e.g. collagenase) family of enzymes from vertebrate origin. Northern and Western blot analysis demonstrated that the metalloproteinase transcript and protein are under a strict developmental program in that both are expressed only in leaf tissue and in a temporal fashion. The physiological function of the metalloproteinase still remains unclear although the data suggest that the enzyme is extracellular and a portion of the mature form of the enzyme is tightly bound to the cell wall.
Pak
J H
JH
Department of Biological Sciences, Bowling Green State University, OH 43403, USA.
Liu
C Y
CY
Huangpu
J
J
Graham
J S
JS
D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D006801:Humans; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D018515:Plant Leaves; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D019105:Random Amplified Polymorphic DNA Technique; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000201:enzymology*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D012333:RNA, Messenger; D008666:Metalloendopeptidases