A prediction of DPP IV/CD26 domain structure from a physico-chemical investigation of dipeptidyl peptidase IV (CD26) from human seminal plasma.
Human DPP IV, isolated from seminal plasma by means of immobilised adenosine deaminase, occurs in different forms which are distinguishable by net charge and native molecular weight. Charge differences arise primarily from different degrees of glycosylation containing various amounts of sialic acid. The majority of DPP IV isolated from total seminal plasma consists of the extracellular part of the protein starting at Gly-31. It is a very stable protein resisting high concentrations of denaturant. Unfolding experiments under reducing conditions are indicative of the existence of at least two domains which function independently. One of these domains is highly stabilised by disulfide bonds. Disruption of the disulfide bonds does not affect the activity, the dimeric state nor the adenosine deaminase binding properties of the protein but renders it more susceptible to proteolysis. The low-angle X-ray scattering spectrum is consistent with a model for a protein containing two subunits, each composed of three domains linked by flexible regions with low average mass. The secondary structure composition, determined by FTIR spectrometry, indicates that 45% of the protein consists of beta-sheets, which is higher than expected from computed secondary structure predictions. Our results provide compelling experimental evidence for the three-domain structure of the extracellular part of DPP IV.
Lambeir
A M
AM
Department of Pharmaceutical Sciences, University of Antwerp (U.I.A.), Wilrijk, Belgium. lambeir@uia.ua.ac.be
Díaz Pereira
J F
JF
Chacón
P
P
Vermeulen
G
G
Heremans
K
K
Devreese
B
B
Van Beeumen
J
J
De Meester
I
I
Scharpé
S
S
D000595:Amino Acid Sequence; D018819:Dipeptidyl Peptidase 4 / Q000737:chemistry* / Q000302:isolation & purification; D006031:Glycosylation; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D008969:Molecular Sequence Data; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012661:Semen / Q000201:enzymology*; D014508:Urea
D006146:Guanidines; D014508:Urea; D018819:Dipeptidyl Peptidase 4; D019791:Guanidine