Myosin cross-reactive antigen of Streptococcus pyogenes M49 encodes a fatty acid double bond hydratase that plays a role in oleic acid detoxification and bacterial virulence.
The myosin cross-reactive antigen (MCRA) protein family is highly conserved among different bacterial species ranging from Gram-positive to Gram-negative bacteria. Besides their ubiquitous occurrence, knowledge about the biochemical and physiological function of MCRA proteins is scarce. Here, we show that MCRA protein from Streptococcus pyogenes M49 is a FAD enzyme, which acts as hydratase on (9Z)- and (12Z)-double bonds of C-16, C-18 non-esterified fatty acids. Products are 10-hydroxy and 10,13-dihydroxy fatty acids. Kinetic analysis suggests that FAD rather stabilizes the active conformation of the enzyme and is not directly involved in catalysis. Analysis of S. pyogenes M49 grown in the presence of either oleic or linoleic acid showed that 10-hydroxy and 10,13-dihydroxy derivatives were the only products. No further metabolism of these hydroxy fatty acids was detected. Deletion of the hydratase gene caused a 2-fold decrease in minimum inhibitory concentration against oleic acid but increased survival of the mutant strain in whole blood. Adherence and internalization properties to human keratinocytes were reduced in comparison with the wild type. Based on these results, we conclude that the previously identified MCRA protein can be classified as a FAD-containing double bond hydratase, within the carbon-oxygen lyase family, that plays a role in virulence of at least S. pyogenes M49.
Volkov
Anton
A
Department for Plant Biochemistry, Albrecht-von-Haller-Institute for Plant Sciences, Georg-August-University, 37077 Göttingen, Germany.
Liavonchanka
Alena
A
Kamneva
Olga
O
Fiedler
Tomas
T
Goebel
Cornelia
C
Kreikemeyer
Bernd
B
Feussner
Ivo
I
D000595:Amino Acid Sequence; D002470:Cell Survival; D003429:Cross Reactions; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006836:Hydro-Lyases / Q000235:genetics* / Q000378:metabolism*; D015603:Keratinocytes / Q000382:microbiology; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009218:Myosins / Q000378:metabolism*; D019301:Oleic Acid / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013290:Streptococcal Infections / Q000378:metabolism* / Q000382:microbiology; D013297:Streptococcus pyogenes / Q000235:genetics / Q000472:pathogenicity*; D014774:Virulence / Q000502:physiology*; D037521:Virulence Factors / Q000235:genetics* / Q000378:metabolism*
D011994:Recombinant Proteins; D037521:Virulence Factors; D005182:Flavin-Adenine Dinucleotide; D019301:Oleic Acid; D009218:Myosins; D006836:Hydro-Lyases; C549330:myosin cross-reactive antigen, Streptococcus pyogenes