Structure and kinetic characterization of human sperm-specific

Structure and kinetic characterization of human
sperm-specific glyceraldehyde-3-phosphate
dehydrogenase, GAPDS
Apirat Chaikuad, Naeem Shafqat, Ruby Al-Mokhtar, Gus Cameron, Anthony
R Clarke, Leo Brady, Udo Oppermann, Jan Frayne, Wyatt W. Yue
To cite this version:
Apirat Chaikuad, Naeem Shafqat, Ruby Al-Mokhtar, Gus Cameron, Anthony R Clarke, et
al.. Structure and kinetic characterization of human sperm-specific glyceraldehyde-3-phosphate
dehydrogenase, GAPDS. Biochemical Journal, Portland Press, 2011, 435 (2), pp.401-409. .
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Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442
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