Watermelon glyoxysomal malate dehydrogenase is sorted to peroxisomes of the methylotrophic yeast, Hansenula polymorpha.
We have studied the fate of the watermelon (Citrullus vulgaris Schrad.) glyoxysomal enzyme, malate dehydrogenase (gMDH), after synthesis in the methylotrophic yeast, Hansenula polymorpha. The gene encoding the precursor form of gMDH (pre-gMDH) was cloned in an H. polymorpha expression vector downstream of the inducible H. polymorpha alcohol oxidase promoter. During methylotrophic growth, pre-gMDH was synthesized and imported into peroxisomes, where it was enzymatically active. The apparent molecular mass of the protein located in H. polymorpha peroxisomes was equal to that of pre-gMDH (41 kDa), indicating that N-terminal processing of the transit peptide had not occurred in the yeast.
van der Klei
I J
IJ
Laboratory for Electron Microscopy, University of Groningen, Haren, The Netherlands.
Faber
K N
KN
Keizer-Gunnink
I
I
Gietl
C
C
Harder
W
W
Veenhuis
M
M
D015153:Blotting, Western; D003001:Cloning, Molecular; D005638:Fruit / Q000201:enzymology*; D008291:Malate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D008830:Microbodies / Q000201:enzymology*; D008854:Microscopy, Electron; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D011498:Protein Precursors; D011994:Recombinant Proteins; D008291:Malate Dehydrogenase