Structural and Kinetic Characterization of the SpeG Spermidine/Spermine N-acetyltransferase from Methicillin-Resistant Staphylococcus aureus USA300

被引:6
作者
Tsimbalyuk, Sofiya [1 ]
Shornikov, Aleksander [2 ]
Srivastava, Parul [1 ]
Le, Van Thi Bich [2 ]
Warren, Imani [2 ]
Khandokar, Yogesh B. B. [1 ]
Kuhn, Misty L. L. [2 ]
Forwood, Jade K. K. [1 ,3 ]
机构
[1] Charles Sturt Univ, Sch Dent & Med Sci, Boorooma St, Wagga Wagga, NSW 2678, Australia
[2] San Francisco State Univ, Deparment Chem & Biochem, San Francisco, CA 94132 USA
[3] Charles Sturt Univ, Sch Biomed Sci, Wagga Wagga, NSW 2678, Australia
基金
美国国家卫生研究院;
关键词
Gcn5-related N-acetyltransferase (GNAT); Staphylococcus aureus; acetylation; spermidine; spermine N-acetyltransferase (SSAT); polyamine; MRSA USA300; CATABOLIC MOBILE ELEMENT; VIBRIO-CHOLERAE; REFINEMENT; POLYAMINES; VIRULENCE; SEQUENCE; FEATURES; CLONE;
D O I
10.3390/cells12141829
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Polyamines are simple yet critical molecules with diverse roles in numerous pathogenic and non-pathogenic organisms. Regulating polyamine concentrations affects the transcription and translation of genes and proteins important for cell growth, stress, and toxicity. One way polyamine concentrations are maintained within the cell is via spermidine/spermine N-acetyltransferases (SSATs) that acetylate intracellular polyamines so they can be exported. The bacterial SpeG enzyme is an SSAT that exhibits a unique dodecameric structure and allosteric site compared to other SSATs that have been previously characterized. While its overall 3D structure is conserved, its presence and role in different bacterial pathogens are inconsistent. For example, not all bacteria have speG encoded in their genomes; in some bacteria, the speG gene is present but has become silenced, and in other bacteria, it has been acquired on mobile genetic elements. The latter is the case for methicillin-resistant Staphylococcus aureus (MRSA) USA300, where it appears to aid pathogenesis. To gain a greater understanding of the structure/function relationship of SpeG from the MRSA USA300 strain (SaSpeG), we determined its X-ray crystal structure in the presence and absence of spermine. Additionally, we showed the oligomeric state of SaSpeG is dynamic, and its homogeneity is affected by polyamines and AcCoA. Enzyme kinetic assays showed that pre-incubation with polyamines significantly affected the positive cooperativity toward spermine and spermidine and the catalytic efficiency of the enzyme. Furthermore, we showed bacterial SpeG enzymes do not have equivalent capabilities to acetylate aminopropyl versus aminbutyl ends of spermidine. Overall, this study provides new insight that will assist in understanding the SpeG enzyme and its role in pathogenic and non-pathogenic bacteria at a molecular level.
引用
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页数:20
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