SAMP14, a novel, acrosomal membrane-associated, glycosylphosphatidylinositol-anchored member of the Ly-6/urokinase-type plasminogen activator receptor superfamily with a role in sperm-egg interaction

Jagathpala Shetty, Michael J. Wolkowicz, Laura C. Digilio, Kenneth L. Klotz, Friederike L. Jayes, Alan B. Diekman, V. Anne Westbrook, Erin M. Farris, Zhonglin Hao, Scott A. Coonrod, Charles J. Flickinger, John C. Herr

Research output: Contribution to journalArticle

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Abstract

We report a new member of the Ly-6/urokinase-type plasminogen activator receptor (uPAR) superfamily of receptors, SAMP14, which is retained on the inner acrosomal membrane of the human spermatozoan following the acrosome reaction and may play a role in fertilization. The SAMP14 sequence predicted a glycosylphosphatidylinositol (GPI)-anchored protein with a signal peptide, a transmembrane domain near the carboxyl terminus, and a putative transamidase cleavage site in the proprotein. Attachment of SAMP14 to the membrane by a lipid anchor was confirmed by its sensitivity to phosphatidylinositol phospholipase C. SAMP14 has a single functional domain similar to the Ly-6 and urokinase plasminogen activator receptor superfamily of proteins, and the gene mapped to 19q13.33, near the PLAUR locus for uPAR at 19q13.2. Northern and dot blotting showed that SAMP14 expression was testis-specific. Indirect immunofluorescence and immunoelectron microscopy with antisera to purified recombinant SAMP14 localized the protein to outer and inner acrosomal membranes as well as the acrosomal matrix of ejaculated human sperm. Acrosome-reacted sperm demonstrated SAMP14 immunofluorescence, indicating its retention on the inner acrosomal membrane following the acrosome reaction. However, SAMP14 localized to the entire sperm when unwashed swim-up sperm from the ejaculate were stained, indicating that some SAMP14 is loosely associated with the plasma membrane. Antibodies against recombinant SAMP14 inhibited both the binding and the fusion of human sperm to zona free hamster eggs, suggesting that SAMP14 may have a role in sperm-egg interaction. SAMP14 represents a GPI-anchored putative receptor in the Ly-6/uPAR family that is exposed on the inner acrosomal membrane after the acrosome reaction.

Original languageEnglish (US)
Pages (from-to)30506-30515
Number of pages10
JournalJournal of Biological Chemistry
Volume278
Issue number33
DOIs
StatePublished - Aug 15 2003

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Urokinase Plasminogen Activator Receptors
Sperm-Ovum Interactions
Glycosylphosphatidylinositols
Spermatozoa
Acrosome Reaction
Membranes
Acrosome
Proteins
Immunoelectron Microscopy
Herpes Zoster
Type C Phospholipases
Cell membranes
Indirect Fluorescent Antibody Technique
Membrane Lipids
Protein Sorting Signals
Phosphatidylinositols
Anchors
Fluorescence Microscopy
Fertilization
Cricetinae

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

SAMP14, a novel, acrosomal membrane-associated, glycosylphosphatidylinositol-anchored member of the Ly-6/urokinase-type plasminogen activator receptor superfamily with a role in sperm-egg interaction. / Shetty, Jagathpala; Wolkowicz, Michael J.; Digilio, Laura C.; Klotz, Kenneth L.; Jayes, Friederike L.; Diekman, Alan B.; Westbrook, V. Anne; Farris, Erin M.; Hao, Zhonglin; Coonrod, Scott A.; Flickinger, Charles J.; Herr, John C.

In: Journal of Biological Chemistry, Vol. 278, No. 33, 15.08.2003, p. 30506-30515.

Research output: Contribution to journalArticle

Shetty, J, Wolkowicz, MJ, Digilio, LC, Klotz, KL, Jayes, FL, Diekman, AB, Westbrook, VA, Farris, EM, Hao, Z, Coonrod, SA, Flickinger, CJ & Herr, JC 2003, 'SAMP14, a novel, acrosomal membrane-associated, glycosylphosphatidylinositol-anchored member of the Ly-6/urokinase-type plasminogen activator receptor superfamily with a role in sperm-egg interaction', Journal of Biological Chemistry, vol. 278, no. 33, pp. 30506-30515. https://doi.org/10.1074/jbc.M301713200
Shetty, Jagathpala ; Wolkowicz, Michael J. ; Digilio, Laura C. ; Klotz, Kenneth L. ; Jayes, Friederike L. ; Diekman, Alan B. ; Westbrook, V. Anne ; Farris, Erin M. ; Hao, Zhonglin ; Coonrod, Scott A. ; Flickinger, Charles J. ; Herr, John C. / SAMP14, a novel, acrosomal membrane-associated, glycosylphosphatidylinositol-anchored member of the Ly-6/urokinase-type plasminogen activator receptor superfamily with a role in sperm-egg interaction. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 33. pp. 30506-30515.
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abstract = "We report a new member of the Ly-6/urokinase-type plasminogen activator receptor (uPAR) superfamily of receptors, SAMP14, which is retained on the inner acrosomal membrane of the human spermatozoan following the acrosome reaction and may play a role in fertilization. The SAMP14 sequence predicted a glycosylphosphatidylinositol (GPI)-anchored protein with a signal peptide, a transmembrane domain near the carboxyl terminus, and a putative transamidase cleavage site in the proprotein. Attachment of SAMP14 to the membrane by a lipid anchor was confirmed by its sensitivity to phosphatidylinositol phospholipase C. SAMP14 has a single functional domain similar to the Ly-6 and urokinase plasminogen activator receptor superfamily of proteins, and the gene mapped to 19q13.33, near the PLAUR locus for uPAR at 19q13.2. Northern and dot blotting showed that SAMP14 expression was testis-specific. Indirect immunofluorescence and immunoelectron microscopy with antisera to purified recombinant SAMP14 localized the protein to outer and inner acrosomal membranes as well as the acrosomal matrix of ejaculated human sperm. Acrosome-reacted sperm demonstrated SAMP14 immunofluorescence, indicating its retention on the inner acrosomal membrane following the acrosome reaction. However, SAMP14 localized to the entire sperm when unwashed swim-up sperm from the ejaculate were stained, indicating that some SAMP14 is loosely associated with the plasma membrane. Antibodies against recombinant SAMP14 inhibited both the binding and the fusion of human sperm to zona free hamster eggs, suggesting that SAMP14 may have a role in sperm-egg interaction. SAMP14 represents a GPI-anchored putative receptor in the Ly-6/uPAR family that is exposed on the inner acrosomal membrane after the acrosome reaction.",
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T1 - SAMP14, a novel, acrosomal membrane-associated, glycosylphosphatidylinositol-anchored member of the Ly-6/urokinase-type plasminogen activator receptor superfamily with a role in sperm-egg interaction

AU - Shetty, Jagathpala

AU - Wolkowicz, Michael J.

AU - Digilio, Laura C.

AU - Klotz, Kenneth L.

AU - Jayes, Friederike L.

AU - Diekman, Alan B.

AU - Westbrook, V. Anne

AU - Farris, Erin M.

AU - Hao, Zhonglin

AU - Coonrod, Scott A.

AU - Flickinger, Charles J.

AU - Herr, John C.

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N2 - We report a new member of the Ly-6/urokinase-type plasminogen activator receptor (uPAR) superfamily of receptors, SAMP14, which is retained on the inner acrosomal membrane of the human spermatozoan following the acrosome reaction and may play a role in fertilization. The SAMP14 sequence predicted a glycosylphosphatidylinositol (GPI)-anchored protein with a signal peptide, a transmembrane domain near the carboxyl terminus, and a putative transamidase cleavage site in the proprotein. Attachment of SAMP14 to the membrane by a lipid anchor was confirmed by its sensitivity to phosphatidylinositol phospholipase C. SAMP14 has a single functional domain similar to the Ly-6 and urokinase plasminogen activator receptor superfamily of proteins, and the gene mapped to 19q13.33, near the PLAUR locus for uPAR at 19q13.2. Northern and dot blotting showed that SAMP14 expression was testis-specific. Indirect immunofluorescence and immunoelectron microscopy with antisera to purified recombinant SAMP14 localized the protein to outer and inner acrosomal membranes as well as the acrosomal matrix of ejaculated human sperm. Acrosome-reacted sperm demonstrated SAMP14 immunofluorescence, indicating its retention on the inner acrosomal membrane following the acrosome reaction. However, SAMP14 localized to the entire sperm when unwashed swim-up sperm from the ejaculate were stained, indicating that some SAMP14 is loosely associated with the plasma membrane. Antibodies against recombinant SAMP14 inhibited both the binding and the fusion of human sperm to zona free hamster eggs, suggesting that SAMP14 may have a role in sperm-egg interaction. SAMP14 represents a GPI-anchored putative receptor in the Ly-6/uPAR family that is exposed on the inner acrosomal membrane after the acrosome reaction.

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