The Chlamydomonas reinhardtii MoCo carrier protein is multimeric and stabilizes molybdopterin cofactor in a molybdate charged form.
In Chlamydomonas reinhardtii, molybdopterin cofactor (MoCo) able to reconstitute active nitrate reductase (NR) with apoenzyme from the Neurospora crassa mutant nit-1 was found mostly bound to a carrier protein (CP). This protein is scarce in the algal free extracts and has been purified 520-fold. MoCoCP is a protein of 64 kDa with subunits of 16.5 kDa and an isoelectric point of 4.5. In contrast to free MoCo, MoCo bound to CP was remarkably protected against inactivation under both aerobic conditions and basic pH. MocoCP transferred active MoCo to apoNR in vitro without addition of molybdate, though reconstituted activity was 20% higher in the presence of molybdate. Incubation with tungstate specifically inhibited MoCoCP activity but had no effect on the activity of free MoCo released from milk xanthine oxidase. MoCoCP did not charge molybdate unless in the presence of N. crassa extracts. Our data support that MoCoCP stabilizes MoCo in an active form charged with molybdate to provide MoCo to apomolybdoenzymes.
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias e Instituto Andaluz de Biotecnología, Universidad de Córdoba, Spain.
C P
CP
Witte
M I
MI
Igeño
R
R
Mendel
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G
Schwarz
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E
Fernández
D000818:Animals; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000187:drug effects / Q000378:metabolism*; D003067:Coenzymes; D008667:Metalloproteins / Q000302:isolation & purification / Q000378:metabolism*; D008982:Molybdenum / Q000737:chemistry / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D011621:Pteridines / Q000302:isolation & purification / Q000378:metabolism*; D017973:Tungsten Compounds / Q000494:pharmacology
D002352:Carrier Proteins; D003067:Coenzymes; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D011621:Pteridines; D017973:Tungsten Compounds; C044659:molybdate; C022261:molybdopterin; C045951:tungstate