SHBG region of the anticoagulant cofactor protein S: secondary structure prediction, circular dichroism spectroscopy, and analysis of naturally occurring mutations.
Protein S (PS) and growth arrest specific factor 6 (GAS6) are vitamin K-dependent proteins with similar structures. They are mosaic proteins possessing a carboxyl-terminal region presenting sequence similarity with plasma sex hormone binding globulin (plasma SHBG), although apparently not involved in steroid binding. The SHBG-like modules have sequence similarity with the G repeats of the chain A of laminin. Laminin G repeats have been reported to contain mainly beta-strands (about 40-50%) but no or little alpha structure by circular dichroism (CD) spectroscopy. Secondary structure predictions carried out in the present work unexpectedly showed a 20 to 27% helices content in the SHBG region of PS/GAS6 (about 100 residues), while plasma SHBG and laminin G repeats had around 10% helices. CD measurements for human PS indicated also that its SHBG region had about 100 residues in alpha-helical structure. These data suggest that the SHBG region of PS/GAS6 on the one hand, and the laminin G repeats and possibly plasma SHBG on the other hand, could present important structural differences. Previously reported polymorphisms and point mutations leading to PS deficiency and thrombophilia have been analyzed with our structural predictions. We found a good agreement between these structural predictions, CD measurements, experimental and clinical data. This information allows us to gain insights into the three-dimensional structure of PS that will be helpful for the design of new experiments and future clinical investigations.
Villoutreix
B O
BO
Department of Clinical Chemistry, Wallenberg Laboratory, Lund University, University Hospital, Malmö, Sweden. bruno.villoutreix@klkemi.mas.lu.se
García de Frutos
P
P
Lövenklev
M
M
Linse
S
S
Fernlund
P
P
Dahlbäck
B
B
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D015935:Complement C4b / Q000378:metabolism; D004252:DNA Mutational Analysis; D000882:Haplorhini; D006649:Histocompatibility Antigens / Q000378:metabolism; D006801:Humans; D007930:Leucine / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011392:Proline / Q000235:genetics; D017293:Protein S / Q000737:chemistry* / Q000235:genetics*; D018455:Protein S Deficiency / Q000235:genetics; D017433:Protein Structure, Secondary; D011817:Rabbits; D051381:Rats; D011987:Receptors, Steroid / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012738:Sex Hormone-Binding Globulin / Q000737:chemistry* / Q000235:genetics*; D012997:Solvents; D013552:Swine
D006649:Histocompatibility Antigens; D017293:Protein S; D011987:Receptors, Steroid; D012738:Sex Hormone-Binding Globulin; D012997:Solvents; D015935:Complement C4b; D011392:Proline; D007930:Leucine