| 1995 |
AKAP4 (Fsc1) encodes the major structural protein of the fibrous sheath in the principal piece of the sperm flagellum; mRNA is expressed exclusively in postmeiotic (round) spermatids and the protein contains 32 cysteine residues and 32 potential phosphorylation sites with no homology to other known cytoskeletal proteins. |
cDNA cloning, Northern blot, in situ hybridization, peptide sequencing |
Biology of reproduction |
High |
7711182
|
| 1997 |
Pro-AKAP4 (pro-AKAP82) is synthesized in the spermatid cell body, transported down the axoneme, proteolytically cleaved to form mature AKAP4 during fibrous sheath assembly, and becomes insoluble (Triton X-100 resistant) in mature sperm; the RII subunit of PKA co-purifies with AKAP4 in the particulate fraction of sperm, consistent with AKAP4 anchoring PKA to the fibrous sheath. |
Immunoelectron microscopy, immunoblotting with phosphatase treatment, Triton X-100 fractionation, antiserum against pro-domain peptide |
Developmental biology |
High |
9441672
|
| 1998 |
AKAP4 (FSC1) contains two distinct RIα tethering domains: domain A (residues 219–232) binds both RIα and RII (dual-specificity), and domain B (residues 335–344) specifically binds RIα via an amphipathic helix; site-directed mutagenesis disrupting the amphipathic helix of domain B abolishes RIα binding. |
Yeast two-hybrid screening of mouse testis cDNA library, GST-fusion in vitro binding assays, deletion analysis, site-directed mutagenesis, helical wheel projection |
The Journal of biological chemistry |
High |
9852104
|
| 1998 |
Human AKAP4 (hAKAP82) is encoded by an X-linked gene (Xp11.2), is highly homologous to mouse AKAP82, and its precursor pro-hAKAP82 binds the type II regulatory subunit of PKA through a domain identical to that of the mouse protein; alternative splicing generates at least two distinct transcripts. |
cDNA cloning, RII overlay assay, immunofluorescence, chromosomal mapping, sequence analysis |
The Journal of biological chemistry |
High |
9822690
|
| 1999 |
Bovine AKAP82 binds the regulatory subunit of PKA and localizes to the entire principal piece of the sperm flagellum, demonstrating conservation of PKA-anchoring function across mammals. |
cDNA cloning, RII binding assay, immunofluorescence, immunoblotting |
Biology of reproduction |
Medium |
10411509
|
| 1999 |
Pro-hAKAP82 and hAKAP82 undergo capacitation-dependent tyrosine phosphorylation in human spermatozoa, but neither the extent of proteolytic processing nor tyrosine phosphorylation of these proteins was found to correlate with differences in sperm motility. |
Immunofluorescence, immunoblotting with phosphotyrosine antibodies |
Molecular human reproduction |
Medium |
10460219
|
| 2002 |
Genetic knockout of Akap4 in mice causes failure of fibrous sheath assembly, loss of progressive sperm motility, and male infertility without reducing sperm numbers; fibrous sheath-associated proteins (including signaling and glycolytic enzymes) are absent or substantially reduced, establishing AKAP4 as an essential scaffold for fibrous sheath organization. |
Gene targeting (knockout mouse), electron microscopy, immunoblotting, motility analysis |
Developmental biology |
High |
12167408
|
| 2004 |
In Akap4-knockout mice, PKA catalytic and regulatory subunits and PI3-kinase lose their normal fibrous sheath subcellular distribution, and PP1γ2 phosphorylation and activity are significantly altered, demonstrating that the fibrous sheath scaffold formed by AKAP4 is required for correct spatial organization and regulation of these kinase/phosphatase signaling components. |
Akap4-knockout mouse, subcellular fractionation, immunoblotting, phosphatase activity assay |
Biology of reproduction |
High |
15385410
|
| 2006 |
The pro domain of AKAP4 keeps the precursor in a diffuse cytoplasmic localization in somatic cells; upon removal of the pro domain, mature AKAP4 adopts a punctate distribution dependent on the microtubular cytoskeleton via two domains (T2a and T2b) homologous to the actin-binding T2-tethering domain of AKAP5, suggesting AKAP4 interacts with microtubules to be transported/incorporated into the fibrous sheath. |
GFP-fusion protein expression in somatic cell lines, immunofluorescence, cytoskeletal drug treatments (microtubule/actin disruption) |
Biology of reproduction |
Medium |
16687648
|
| 2005 |
The Akap4 gene produces two alternatively spliced transcripts (Akap82 and Fsc1) differing only in their 5′ UTRs; only the Akap82 transcript is loaded onto polyribosomes and translated in spermatids, while the Fsc1 transcript is not polysomal despite both being deadenylated during spermiogenesis. |
Polyribosome fractionation, RT-PCR, reporter assays in vivo and in vitro |
Molecular reproduction and development |
Medium |
15685631
|
| 2012 |
ERK1/2 phosphorylates proAKAP4 on Thr265 in human spermatozoa in vitro and in vivo; phosphorylation by ERK1/2 (downstream of PKC) causes relocalization of proAKAP4 from the principal piece to the Golgi/mid-piece in a T265-dependent manner; cAMP/PKA attenuates PKC-dependent ERK1/2 activation only in the presence of proAKAP4, and disruption of PKA-RII/AKAP binding (St-HT31) abolishes this inhibitory cross-talk, establishing proAKAP4 as a molecular switch between cAMP/PKA and PKC/ERK1/2 pathways. |
In vitro kinase assay, site-directed mutagenesis (T265A), GFP-proAKAP4 transfection in HEK293T, immunofluorescence, phorbol ester stimulation, PKA-AKAP disrupting peptide St-HT31 |
Scientific reports |
High |
27901058
|
| 2012 |
Porcine oviductal DMBT1 induces tyrosine phosphorylation of proAKAP4 (but not the mature ~80 kDa AKAP4) localized to periacrosomal membranes, independently of calcium, bicarbonate, cAMP analogs, PKA inhibitors, or PKC inductor, placing proAKAP4 phosphorylation as an early step in the DMBT1-triggered signal transduction pathway controlling sperm selection. |
Mass spectrometry, immunoprecipitation, immunofluorescence, subcellular fractionation, pharmacological inhibitor panel |
Reproduction (Cambridge, England) |
Medium |
22457434
|
| 2017 |
AKAP4 knockdown in ovarian cancer cells leads to PKA protein degradation that is rescued by the proteasome inhibitor MG-132, increased ROS and DNA damage, and cell cycle arrest rescued by the ROS quencher NAC, demonstrating that AKAP4 maintains PKA stability and acts through the PKA-CREB signaling axis in cancer cells. |
siRNA knockdown, proteasome inhibitor rescue (MG-132), NAC treatment, immunoblotting, flow cytometry, SCID mouse xenograft |
Oncotarget |
Medium |
28881798
|
| 2021 |
A hemizygous missense variant (c.1285C>T, p.R429C) in AKAP4 reduces AKAP4 protein expression and its interaction with QRICH2 (demonstrated by co-immunoprecipitation and co-localization), resulting in decreased QRICH2 protein in spermatozoa and dysplastic fibrous sheath leading to male infertility. |
Whole-exome sequencing, co-immunoprecipitation, immunofluorescence, HEK-293T cell expression |
Human molecular genetics |
Medium |
34415320
|
| 2021 |
A missense mutation p.S152P in the pro-domain of AKAP4 causes protein accumulation in the cytoplasm of COS-7 cells (failure to traffic correctly), abolishes mature AKAP4 in spermatozoa, increases ROS/apoptotic markers in GC2-spd cells, decreases PKA/PI3K signaling activity, and disrupts the interaction between AKAP4 and RNASET2. |
Sanger sequencing, COS-7 cell transfection with GFP-fusion constructs, immunofluorescence, co-immunoprecipitation, flow cytometry, oxidative stress assays |
Molecular reproduction and development |
Medium |
34409659
|
| 2024 |
AKAP4 is a target of SUMO1 modification in porcine sperm under cryopreservation/oxidative stress; SUMO1 modification level increases while AKAP4 protein level decreases under these conditions, but inhibition of SUMO1 modification does not affect AKAP4 degradation, indicating SUMO1 is not involved in AKAP4 proteolysis. However, global inhibition of sperm protein SUMO1 modification reduces sperm motility, ATP content, and DNA integrity. |
LC-MS/MS identification of SUMO-modified proteins, SUMO inhibitor treatment, immunoblotting, sperm motility and ATP assays |
Animal reproduction science |
Low |
39765132
|
| 2020 |
Phosphoproteomics of mouse sperm capacitation identified phosphorylation of AKAP4 at Y156 (increased) and Y811 (decreased) during capacitation, with overall upregulated phospho-AKAP4 trends detected by western blot, indicating AKAP4 phosphorylation is dynamically regulated during capacitation. |
Label-free quantitative phosphoproteomics, western blotting, IPA pathway analysis |
International journal of molecular sciences |
Low |
33023073
|
| 2025 |
SPAG6 modulates testicular AKAP4 levels in mice; in compound Spag6/Spag6l mutants with disorganized fibrous sheath, AKAP4 levels are altered, suggesting SPAG6 acts upstream of AKAP4 in the pathway controlling fibrous sheath assembly. |
Double-knockout mouse model, immunoblotting, histological and ultrastructural analysis |
bioRxivpreprint |
Low |
bio_10.1101_2025.07.18.665465
|