Purification and functional characterization of diadenosine 5',5‴-P(1),P(4)-tetraphosphate phosphorylases from Mycobacterium smegmatis and Mycobacterium avium.
We recently demonstrated that the Rv2613c protein from Mycobacterium tuberculosis H37Rv is a novel diadenosine 5',5‴-P(1),P(4)-tetraphosphate (Ap4A) phosphorylase (MtAPA) that forms a tetramer. Mycobacterium avium and Mycobacterium smegmatis express proteins named MAV_3489 and MSMEG_2932, respectively, that are homologous to MtAPA. Here we showed that the MAV_3489 and MSMEG_2932 proteins possess Ap4A phosphorylase activity and enzymatic properties similar to those of MtAPA. Furthermore, gel-filtration column chromatography revealed that MAV_3489 and MSMEG_2932 assembled into homotetramers in solution, indicating that they may also form unique Ap4A-binding sites composed of tetramers.
Department of Quality Assurance & Radiological Protection, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjyuku-ku, Tokyo 162-8640, Japan.
Naoko
N
Honda
Department of Bacteriology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan.
Hyun
H
Kim
Department of Bacteriology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan.
Emiko
E
Rimbara
Department of Quality Assurance & Radiological Protection, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjyuku-ku, Tokyo 162-8640, Japan.
Atsushi
A
Kato
Department of Bacteriology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan.
Keigo
K
Shibayama
Department of Bacteriology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan. Electronic address: mshige@nih.go.jp.
Shigetarou
S
Mori
D000595:Amino Acid Sequence; D000818:Animals; D002413:Cations, Divalent / Q000378:metabolism; D003001:Cloning, Molecular; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D009165:Mycobacterium Infections, Nontuberculous / Q000382:microbiology; D009162:Mycobacterium avium / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D020102:Mycobacterium smegmatis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity; D014379:Tuberculosis, Avian / Q000382:microbiology
D002413:Cations, Divalent; D008670:Metals; D011994:Recombinant Proteins; D009713:Nucleotidyltransferases; C044740:diadenosine 5,5'''-P(1),P(4)-tetraphosphate alpha,beta-phosphorylase
Drug target; Kinetics; Nontuberculous mycobacteria; Nucleotide phosphorylase